CN108004187A - A kind of lactobacillus gasseri and its application for being used to prepare vagina antibacterial medicines - Google Patents
A kind of lactobacillus gasseri and its application for being used to prepare vagina antibacterial medicines Download PDFInfo
- Publication number
- CN108004187A CN108004187A CN201810026176.2A CN201810026176A CN108004187A CN 108004187 A CN108004187 A CN 108004187A CN 201810026176 A CN201810026176 A CN 201810026176A CN 108004187 A CN108004187 A CN 108004187A
- Authority
- CN
- China
- Prior art keywords
- lactobacillus gasseri
- lactobacillus
- vaginal
- vagina
- gasseri
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 241000186606 Lactobacillus gasseri Species 0.000 title claims abstract description 98
- 210000001215 vagina Anatomy 0.000 title claims abstract description 40
- 239000003814 drug Substances 0.000 title claims abstract description 17
- 230000000844 anti-bacterial effect Effects 0.000 title claims abstract description 14
- 229940079593 drug Drugs 0.000 title claims abstract description 10
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims abstract description 24
- 238000004519 manufacturing process Methods 0.000 claims abstract description 13
- 244000052616 bacterial pathogen Species 0.000 claims abstract description 9
- 210000002919 epithelial cell Anatomy 0.000 claims abstract description 8
- 239000002068 microbial inoculum Substances 0.000 claims abstract description 7
- 208000013464 vaginal disease Diseases 0.000 claims abstract description 6
- 241000894006 Bacteria Species 0.000 claims description 45
- 241000207201 Gardnerella vaginalis Species 0.000 claims description 6
- 241000607142 Salmonella Species 0.000 claims description 6
- 208000015181 infectious disease Diseases 0.000 claims description 6
- 241000222122 Candida albicans Species 0.000 claims description 4
- 241000588724 Escherichia coli Species 0.000 claims description 4
- 241000191967 Staphylococcus aureus Species 0.000 claims description 4
- 229940095731 candida albicans Drugs 0.000 claims description 4
- 230000001717 pathogenic effect Effects 0.000 claims description 4
- 206010046914 Vaginal infection Diseases 0.000 claims description 3
- 201000008100 Vaginitis Diseases 0.000 claims description 3
- 241000589517 Pseudomonas aeruginosa Species 0.000 claims description 2
- 201000007096 Vulvovaginal Candidiasis Diseases 0.000 claims description 2
- 230000006806 disease prevention Effects 0.000 claims description 2
- 239000000284 extract Substances 0.000 claims description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 claims 1
- 230000001580 bacterial effect Effects 0.000 abstract description 32
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 abstract description 20
- 235000014655 lactic acid Nutrition 0.000 abstract description 10
- 239000004310 lactic acid Substances 0.000 abstract description 10
- 230000000694 effects Effects 0.000 abstract description 7
- 208000004926 Bacterial Vaginosis Diseases 0.000 abstract description 5
- 208000037009 Vaginitis bacterial Diseases 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 4
- 244000005700 microbiome Species 0.000 abstract description 3
- 231100000252 nontoxic Toxicity 0.000 abstract description 3
- 230000003000 nontoxic effect Effects 0.000 abstract description 3
- 230000004956 cell adhesive effect Effects 0.000 abstract description 2
- 201000010099 disease Diseases 0.000 abstract description 2
- 241000186660 Lactobacillus Species 0.000 description 27
- 238000012360 testing method Methods 0.000 description 27
- 229940039696 lactobacillus Drugs 0.000 description 26
- 238000002474 experimental method Methods 0.000 description 22
- 238000000034 method Methods 0.000 description 18
- 241000186673 Lactobacillus delbrueckii Species 0.000 description 15
- 239000012530 fluid Substances 0.000 description 14
- 229920001817 Agar Polymers 0.000 description 9
- 241001465754 Metazoa Species 0.000 description 9
- 239000008272 agar Substances 0.000 description 9
- 239000001963 growth medium Substances 0.000 description 9
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 8
- 230000005764 inhibitory process Effects 0.000 description 7
- 239000002609 medium Substances 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 238000012408 PCR amplification Methods 0.000 description 6
- 238000002513 implantation Methods 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- 230000028327 secretion Effects 0.000 description 6
- 241000196324 Embryophyta Species 0.000 description 5
- 239000012634 fragment Substances 0.000 description 5
- 238000004321 preservation Methods 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- 238000012163 sequencing technique Methods 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 229920000742 Cotton Polymers 0.000 description 4
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 4
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 230000001408 fungistatic effect Effects 0.000 description 4
- 229910052742 iron Inorganic materials 0.000 description 4
- 229930027917 kanamycin Natural products 0.000 description 4
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical class O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000005259 measurement Methods 0.000 description 4
- 239000002207 metabolite Substances 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 3
- 241000193830 Bacillus <bacterium> Species 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 3
- SIKJAQJRHWYJAI-UHFFFAOYSA-N Indole Chemical compound C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 3
- JVTAAEKCZFNVCJ-UHFFFAOYSA-M Lactate Chemical compound CC(O)C([O-])=O JVTAAEKCZFNVCJ-UHFFFAOYSA-M 0.000 description 3
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 3
- 239000002253 acid Substances 0.000 description 3
- 239000000853 adhesive Substances 0.000 description 3
- 230000001070 adhesive effect Effects 0.000 description 3
- 229940038195 amoxicillin / clavulanate Drugs 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- 238000002829 antibacterial sensitivity test Methods 0.000 description 3
- 210000000481 breast Anatomy 0.000 description 3
- NWFNSTOSIVLCJA-UHFFFAOYSA-L copper;diacetate;hydrate Chemical compound O.[Cu+2].CC([O-])=O.CC([O-])=O NWFNSTOSIVLCJA-UHFFFAOYSA-L 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 238000004043 dyeing Methods 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000011081 inoculation Methods 0.000 description 3
- 235000013372 meat Nutrition 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 235000014347 soups Nutrition 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 description 3
- 231100000820 toxicity test Toxicity 0.000 description 3
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 2
- 108010001478 Bacitracin Proteins 0.000 description 2
- 102000004031 Carboxy-Lyases Human genes 0.000 description 2
- 108090000489 Carboxy-Lyases Proteins 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 241000207202 Gardnerella Species 0.000 description 2
- 108010010803 Gelatin Proteins 0.000 description 2
- 229930182566 Gentamicin Natural products 0.000 description 2
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 2
- 101001076687 Lactobacillus gasseri Inulosucrase Proteins 0.000 description 2
- 101100346764 Mus musculus Mtln gene Proteins 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- 208000005374 Poisoning Diseases 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000004098 Tetracycline Substances 0.000 description 2
- 108010046334 Urease Proteins 0.000 description 2
- 238000009651 Voges-Proskauer test Methods 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 238000009635 antibiotic susceptibility testing Methods 0.000 description 2
- 229960004099 azithromycin Drugs 0.000 description 2
- MQTOSJVFKKJCRP-BICOPXKESA-N azithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)N(C)C[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 MQTOSJVFKKJCRP-BICOPXKESA-N 0.000 description 2
- 229960003071 bacitracin Drugs 0.000 description 2
- 229930184125 bacitracin Natural products 0.000 description 2
- CLKOFPXJLQSYAH-ABRJDSQDSA-N bacitracin A Chemical compound C1SC([C@@H](N)[C@@H](C)CC)=N[C@@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]1C(=O)N[C@H](CCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](CC=2C=CC=CC=2)C(=O)N[C@@H](CC=2N=CNC=2)C(=O)N[C@H](CC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)NCCCC1 CLKOFPXJLQSYAH-ABRJDSQDSA-N 0.000 description 2
- 229960004755 ceftriaxone Drugs 0.000 description 2
- VAAUVRVFOQPIGI-SPQHTLEESA-N ceftriaxone Chemical compound S([C@@H]1[C@@H](C(N1C=1C(O)=O)=O)NC(=O)\C(=N/OC)C=2N=C(N)SC=2)CC=1CSC1=NC(=O)C(=O)NN1C VAAUVRVFOQPIGI-SPQHTLEESA-N 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 210000003756 cervix mucus Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 229960002227 clindamycin Drugs 0.000 description 2
- KDLRVYVGXIQJDK-AWPVFWJPSA-N clindamycin Chemical compound CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@H](C)Cl)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 KDLRVYVGXIQJDK-AWPVFWJPSA-N 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 150000002016 disaccharides Chemical class 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229960003276 erythromycin Drugs 0.000 description 2
- XHCADAYNFIFUHF-TVKJYDDYSA-N esculin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC(C(=C1)O)=CC2=C1OC(=O)C=C2 XHCADAYNFIFUHF-TVKJYDDYSA-N 0.000 description 2
- 239000008273 gelatin Substances 0.000 description 2
- 229920000159 gelatin Polymers 0.000 description 2
- 235000019322 gelatine Nutrition 0.000 description 2
- 235000011852 gelatine desserts Nutrition 0.000 description 2
- 229960002518 gentamicin Drugs 0.000 description 2
- 239000008187 granular material Substances 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 2
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- DMJNNHOOLUXYBV-PQTSNVLCSA-N meropenem Chemical compound C=1([C@H](C)[C@@H]2[C@H](C(N2C=1C(O)=O)=O)[C@H](O)C)S[C@@H]1CN[C@H](C(=O)N(C)C)C1 DMJNNHOOLUXYBV-PQTSNVLCSA-N 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- CEQFOVLGLXCDCX-WUKNDPDISA-N methyl red Chemical compound C1=CC(N(C)C)=CC=C1\N=N\C1=CC=CC=C1C(O)=O CEQFOVLGLXCDCX-WUKNDPDISA-N 0.000 description 2
- 238000000386 microscopy Methods 0.000 description 2
- 235000013336 milk Nutrition 0.000 description 2
- 239000008267 milk Substances 0.000 description 2
- 210000004080 milk Anatomy 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- UWYHMGVUTGAWSP-JKIFEVAISA-N oxacillin Chemical compound N([C@@H]1C(N2[C@H](C(C)(C)S[C@@H]21)C(O)=O)=O)C(=O)C1=C(C)ON=C1C1=CC=CC=C1 UWYHMGVUTGAWSP-JKIFEVAISA-N 0.000 description 2
- 229960001019 oxacillin Drugs 0.000 description 2
- 229940056360 penicillin g Drugs 0.000 description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 230000035479 physiological effects, processes and functions Effects 0.000 description 2
- 229960002292 piperacillin Drugs 0.000 description 2
- IVBHGBMCVLDMKU-GXNBUGAJSA-N piperacillin Chemical compound O=C1C(=O)N(CC)CCN1C(=O)N[C@H](C=1C=CC=CC=1)C(=O)N[C@@H]1C(=O)N2[C@@H](C(O)=O)C(C)(C)S[C@@H]21 IVBHGBMCVLDMKU-GXNBUGAJSA-N 0.000 description 2
- 231100000572 poisoning Toxicity 0.000 description 2
- 230000000607 poisoning effect Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 230000004224 protection Effects 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 229960002180 tetracycline Drugs 0.000 description 2
- 229930101283 tetracycline Natural products 0.000 description 2
- 235000019364 tetracycline Nutrition 0.000 description 2
- 150000003522 tetracyclines Chemical class 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 230000003442 weekly effect Effects 0.000 description 2
- 235000010585 Ammi visnaga Nutrition 0.000 description 1
- 244000153158 Ammi visnaga Species 0.000 description 1
- 241000282693 Cercopithecidae Species 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- HZZVJAQRINQKSD-UHFFFAOYSA-N Clavulanic acid Natural products OC(=O)C1C(=CCO)OC2CC(=O)N21 HZZVJAQRINQKSD-UHFFFAOYSA-N 0.000 description 1
- 208000003322 Coinfection Diseases 0.000 description 1
- 229930182843 D-Lactic acid Natural products 0.000 description 1
- JVTAAEKCZFNVCJ-UWTATZPHSA-N D-lactic acid Chemical compound C[C@@H](O)C(O)=O JVTAAEKCZFNVCJ-UWTATZPHSA-N 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 208000036649 Dysbacteriosis Diseases 0.000 description 1
- 208000027244 Dysbiosis Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- YCKRFDGAMUMZLT-UHFFFAOYSA-N Fluorine atom Chemical compound [F] YCKRFDGAMUMZLT-UHFFFAOYSA-N 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 206010018612 Gonorrhoea Diseases 0.000 description 1
- 238000012449 Kunming mouse Methods 0.000 description 1
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 description 1
- 241000218492 Lactobacillus crispatus Species 0.000 description 1
- 241001561398 Lactobacillus jensenii Species 0.000 description 1
- 241000186783 Lactobacillus vaginalis Species 0.000 description 1
- 241000736262 Microbiota Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- 241000288906 Primates Species 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 208000007074 Trichomonas Vaginitis Diseases 0.000 description 1
- 208000025206 Trichomonas vaginitis urogenital infection Diseases 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 241000607598 Vibrio Species 0.000 description 1
- 206010047400 Vibrio infections Diseases 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000008186 active pharmaceutical agent Substances 0.000 description 1
- 239000005030 aluminium foil Substances 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 229960003022 amoxicillin Drugs 0.000 description 1
- LSQZJLSUYDQPKJ-NJBDSQKTSA-N amoxicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=C(O)C=C1 LSQZJLSUYDQPKJ-NJBDSQKTSA-N 0.000 description 1
- 230000003266 anti-allergic effect Effects 0.000 description 1
- 230000001775 anti-pathogenic effect Effects 0.000 description 1
- 239000004599 antimicrobial Substances 0.000 description 1
- 239000012298 atmosphere Substances 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 238000005842 biochemical reaction Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 230000006037 cell lysis Effects 0.000 description 1
- 239000013553 cell monolayer Substances 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000009642 citrate test Methods 0.000 description 1
- 229960003324 clavulanic acid Drugs 0.000 description 1
- HZZVJAQRINQKSD-PBFISZAISA-N clavulanic acid Chemical compound OC(=O)[C@H]1C(=C/CO)/O[C@@H]2CC(=O)N21 HZZVJAQRINQKSD-PBFISZAISA-N 0.000 description 1
- 230000003749 cleanliness Effects 0.000 description 1
- 238000007621 cluster analysis Methods 0.000 description 1
- 230000019771 cognition Effects 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 229940022769 d- lactic acid Drugs 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 210000000981 epithelium Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 235000015110 jellies Nutrition 0.000 description 1
- 239000008274 jelly Substances 0.000 description 1
- 238000009533 lab test Methods 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000011177 media preparation Methods 0.000 description 1
- 230000005906 menstruation Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000000394 mitotic effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000003097 mucus Anatomy 0.000 description 1
- 239000002547 new drug Substances 0.000 description 1
- 230000027758 ovulation cycle Effects 0.000 description 1
- FWFGVMYFCODZRD-UHFFFAOYSA-N oxidanium;hydrogen sulfate Chemical compound O.OS(O)(=O)=O FWFGVMYFCODZRD-UHFFFAOYSA-N 0.000 description 1
- LSQZJLSUYDQPKJ-UHFFFAOYSA-N p-Hydroxyampicillin Natural products O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)C(N)C1=CC=C(O)C=C1 LSQZJLSUYDQPKJ-UHFFFAOYSA-N 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 235000019371 penicillin G benzathine Nutrition 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 150000002978 peroxides Chemical class 0.000 description 1
- 230000003285 pharmacodynamic effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000012113 quantitative test Methods 0.000 description 1
- 230000003716 rejuvenation Effects 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 230000000452 restraining effect Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- PRWXGRGLHYDWPS-UHFFFAOYSA-L sodium malonate Chemical compound [Na+].[Na+].[O-]C(=O)CC([O-])=O PRWXGRGLHYDWPS-UHFFFAOYSA-L 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 238000013112 stability test Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000041 toxicology testing Toxicity 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 208000000143 urethritis Diseases 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention provides a kind of lactobacillus gasseri and its applications of vagina antibacterial medicines is used to prepare, which is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, its deposit number is CGMCC No.14109.Study it and be metabolized performance discovery, relatively common lactobacillus gasseri, the bacterial strain possesses stronger lactic acid and hydrogen peroxide manufacture ability, very strong to the bacteriostasis of pathogenic bacteria, while has prominent vaginal epithelial cell adhesive capacity.The new property found based on more than, present invention determine that preparing the new application of vagina antibacterial medicines using it, it can be achieved that the treatment of various bacterial vaginal disease.Using microbial inoculum of the present invention to bacterial vaginosis significant effect, and it is safe and non-toxic, stability is good, can preserve for a long time, further relate to its purposes in preventing and/or treating the medicine of gynecological disease.
Description
Technical field
The present invention relates to microbial technology field, adjusting and bacterial vaginosis disease further to microecology in vaginas environment
The treatment of disease, and in particular to a kind of lactobacillus gasseri and its application for being used to prepare vagina antibacterial medicines.
Background technology
For healthy women intravaginal there are multiple-microorganism, they constitute mutually restriction, mutually association between host, environment
Adjust, the microecology in vaginas system of dynamic equilibrium.The vaginal flora of healthy women is mainly made of lactobacillus, including plant breast bar
Bacterium, Lactobacillus Jensenii, lactobacillus gasseri, Lactobacillus crispatus, Lactobacillus vaginalis, lactobacillus gasseri etc..Lactobacillus under normal circumstances
It can play a protective role to vagina, vagina is produced when the lactobacillus disorder of microecology in vaginas can cause pathogenic bacteria to be invaded
It is scorching.
Bacterial vaginosis BV (BV) is due to vaginal dysbacteriosis, and the lactobacillus of host itself reduces and causes it
A large amount of numerous plants of his conditionity pathogenic microorganism such as Gardnerella, various anaerobic bacterias, bending vibrios etc., usual BV be actually with
A kind of mixed infection based on bacterium protein of Gardnerella vaginalis.Using antibiosis extract for treating, can respite BV symptom, but also reduced this
Lactobacillus further reduce, aggravate microecology in vaginas imbalance so that BV recurs repeatedly.How Control in recurring, thoroughly radical cure
Bacterial vaginosis BV is the thorny problem of ob-gyn's urgent need to resolve.
Research shows：Produce H2O2Lactobacillus with lactic acid is the dominant bacteria of healthy women intravaginal, is that protection vagina is exempted from
An important factor for by pathogenic infection, the acid and some antimicrobial agents that in addition lactobacillus metabolism produces can also effectively inhibit
The numerous plant of growth of other bacteriums.
Healthy women intravaginal has individual difference, and anti-pathogenic bacteria energy between each strain of lactobacillus there are a variety of lactobacillus
Force difference is different obvious., it is necessary to consider the species of lactobacillus when selecting lactobacillus probiotics, it produces acid, production H2O2Ability, and with
The ability of vaginal epithelial cell adhesion, can wherein lactobacillus successfully be colonized in vagina, be the basis of lactobacillus continuous action,
It is the key factor that lactobacillus plays curative effect.It is not actually that China's woman vagina is excellent that market, which has product " Lactobacillus delbrueckii ", at present
Gesture flora, colonization ability is poor, can not maintain stable viable bacteria content, it is impossible to meet the needs of gynecological clinic.
Lactobacillus gasseri (Lactobacillus gasseri), is lactobacillus, is one of Body normal flora, extensively
It is distributed in human body intestinal canal, is also distributed in vagina.In the prior art, lactobacillus gasseri is mainly used for stimulating immune thin
Intracrine antiallergy relevant cell hormone；In addition, also intestinal flora feature can be improved by itself flora advantage, thus for normal
For advising bacterial strain, lactobacillus gasseri does not have specific bacteriostasis in the category of cognition of the prior art.
The content of the invention
The present invention is intended to provide a kind of specific lactobacillus gasseri, does not have to solve conventional Grignard lactobacillus in the prior art
There is the technical problem of vaginal pathogenic rejection ability.
Another technical problem to be solved by the present invention is that conventional Grignard lactobacillus production hydrogen peroxide manufacture ability is relatively low.
The invention solves another technical problem be conventional Grignard lactobacillus lactic acid production it is relatively low.
The invention solves another technical problem be in the prior art conventional Grignard lactobacillus colonizing in vagina, deposit
Work is ineffective.
The invention solves another technical problem be in the prior art be directed to vaginal pathogenic microbial inoculum class antibacterial medicines
Therapeutic effect is bad.
To realize above technical purpose, the present invention uses following technical scheme：
A kind of separated lactobacillus gasseri (Lactobacillus gasseri), it is RD-0046 to be numbered in of the invention
(lactobacillus gasseri), is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, its deposit number is
CGMCC No.14109。
Above-mentioned lactobacillus gasseri (Lactobacillus gasseri) RD-0046 is China's Healthy women of child-bearing age vagina point
Screening is got in secretion, is preserved in on May 10th, 2017 in China Committee for Culture Collection of Microorganisms's commonly micro- life
Thing center (abbreviation CGMCC), depositary institution address are：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the micro- life of the Chinese Academy of Sciences
Thing research institute, preservation registration number are CGMCC No.14109, and the Classification And Nomenclature of the bacterial strain is lactobacillus gasseri
(Lactobacillus gasseri)。
Above-mentioned separated lactobacillus gasseri RD-0046, is sequenced using 16SrDNA, with grignard in GenBank databases
Lactobacillus base sequence homology score value is more than 98%.
On the basis of above technical scheme, it is used to prepare vagina invention further provides above-mentioned lactobacillus gasseri and causes
The application of germ antibacterial medicines.
Preferably, the medicine is microbial inoculum, which colonizes and survives on vaginal epithelial cell in vaginal environment.
Preferably, the medicine is microbial inoculum, which is metabolized production H in vaginal environment2O2。
Preferably, the pathogenic bacteria are Gardnerella vaginalis.
Preferably, the pathogenic bacteria are atropic ripple bacterium.
Preferably, the pathogenic bacteria are Candida albicans, staphylococcus aureus, escherichia coli, the false list of verdigris
Born of the same parents bacterium or salmonella etc..
Meanwhile invention further provides above-mentioned lactobacillus gasseri to be used to prepare vaginal disease prevention or medicine
Using.
Preferably, the vaginal disease is vulvovaginal candidiasis, trichomonas vaginitis, senile vagina
Scorching, non-specific vagina infection or Combination vagina infection.
The isolated first one plant of lactobacillus gasseri of the present invention, studies its and is metabolized performance and find that the bacterial strain possesses excellent
Lactic acid production capacity and production hydrogen peroxide energy, it is very strong to the bacteriostasis of pathogenic bacteria, while have the prominent vagina epithelium thin
Born of the same parents' adhesive capacity.The new property found based on more than, present invention determine that using it the new application of vagina antibacterial medicines is prepared, it is real
The treatment of various bacterial vaginal disease is showed.
The beneficial effect produced using above-mentioned technical proposal is：(1) lactobacillus gasseri RD-0046 bacterial strains of the invention can
To preserve for a long time, and bacterial vaginosis BV and various vagina infections are resisted, including candida albicans vaginitis, gonorrhoea, virus
Property vaginitis, and urethral infection etc..(2) bacterial strain of the invention is directly collected in healthy human body, has actively stable life
Thing characteristic, without domestication and rejuvenation technique, is directly entered preparation process.(3) bacterial strain of the invention, which has, suppresses to add moral
Receive Salmonella, suppress Candida albicans effect, with it is commercially available control bacterium compared with, advantageous vaginal epithelial cell adhesive force,
With the advantageous ability being colonized in primate vagina.
Brief description of the drawings
Fig. 1 is the full face of lactobacillus gasseri RD-0046 colonial morphologies in the specific embodiment of the invention；
Fig. 2 is the gram stain microscopy photo of lactobacillus gasseri RD-0046 in the specific embodiment of the invention；
Fig. 3 is the 16SrDNA gene PCR amplified productions of lactobacillus gasseri RD-0046 in the specific embodiment of the invention
Electrophoretogram；
Fig. 4 is the lactate detection collection of illustrative plates of lactobacillus gasseri RD-0046 in the specific embodiment of the invention；
Fig. 5 is lactobacillus gasseri RD-046 hydrogen peroxide colour developing 5min and 10min pictures in the specific embodiment of the invention；
Fig. 6 be in the specific embodiment of the invention lactobacillus gasseri RD-0046 to kanamycins, fleraxacin and A Moxi
The antibiotic susceptibility test bacterium colony figure of woods/clavulanic acid；
Fig. 7 be in the specific embodiment of the invention lactobacillus gasseri RD-0046 (left side) and Lactobacillus delbrueckii (right side) to vagina
The fungistatic effect photo of Gardnerella；
Fig. 8 be in the specific embodiment of the invention lactobacillus gasseri RD-0046 (left side) and Lactobacillus delbrueckii (right side) to sramana
The fungistatic effect photo of Salmonella；
Fig. 9 be in the specific embodiment of the invention lactobacillus gasseri RD-0046 (left side) and Lactobacillus delbrueckii (right side) to verdigris
The fungistatic effect photo of pseudomonad；
Figure 10 is that lactobacillus gasseri RD-0046 is colonized machin vagina after machin vagina in the specific embodiment of the invention
The electrophoretogram of microbiota part bacterial strain 16SrDNA fragment pcr amplification products；
Figure 11 is the scanning electron microscope (SEM) photograph of lactobacillus gasseri RD-0046 of the present invention.
Embodiment
The embodiment of the present invention will be described in detail below.In order to avoid excessive unnecessary details,
It will not be described in detail in following embodiments to belonging to known structure or function.In addition to being defined, institute in following embodiments
Technical and scientific term has the identical meanings being commonly understood by with those skilled in the art of the invention.
Test reagent consumptive material used, is routine biochemistry reagent unless otherwise specified in following embodiments；The experiment
Method, is conventional method unless otherwise specified；Quantitative test in following embodiments, is respectively provided with three repeated experiments, as a result
It is averaged；% in following embodiments, is mass percentage unless otherwise instructed.
Used Bacteria Culture based component and preparation method are as follows in following embodiments：
1st, meat soup solid medium (MRS) is prepared：
(1) by agar powder wiring solution-forming, 1.5g/100ml deionized waters；
(2) MRS culture medium 17.91g/100ml agar solutions are added, are mixed；
(3) autoclave, 1.0MPa steam sterilizings 20 minutes are put into；
(4) treat that culture medium temperature is cooled to room temperature and pour into culture dish, it is a according to culture dish size about 10ml/ or 20ml/；
(5) in being operated in super-clean bench.Into agar shape after cooling, mark culture medium title and prepare the date, be put in 4 DEG C of refrigerators
It is stand-by.
2nd, meat soup fluid nutrient medium (MRS) is prepared：
(1) MRS culture mediums are added into deionized water, ratio is 17.9l g/100ml；
(2) autoclave, 1.0MPa steam sterilizings 20 minutes are put into；
(3) take out, dispense into EP pipes, each 1.0ml after pot no pressure subject to sterilization.Mark culture medium title and prepare day
Phase, it is stand-by to be put in 4 DEG C of refrigerators.
3rd, hydrogen peroxide (H2O2) identification culture medium preparation：
(1) with meat soup solid medium (MRS) preparation steps (1) to (4)；
(2) take out after pot no pressure subject to sterilization, slightly cool down, but still it is (dense eventually in adding TMB in super-clean bench when being liquid condition
Spend 0.25mg/m1), HRP (final concentration 0.01mg/m1), mix；
(3) pour into culture dish after culture medium temperature is down to about 45 DEG C, into agar shape after cooling, mark culture medium title and
The date is prepared, it is stand-by to be put in 4 DEG C of refrigerators.
Embodiment 1 (separation of lactobacillus gasseri RD-0046 floras and inoculation, purifying, Zengjing Granule)
1st, the separation and inoculation of lactobacillus gasseri RD-0046 floras：Subject's vaginal side is gathered with two sterile cotton swabs
Secretion on wall, 24 it is interior when small be inoculated in the culture dish equipped with prepared MRS culture mediums with various concentrations, and mark letter
Breath, culture dish is placed in anaerobic jar, and be put into CO2Aerogenesis bag, is placed in 37 DEG C of incubators, is incubated more than 48h.
2nd, the purifying of lactobacillus gasseri bacterial strain RD-0046, Zengjing Granule：According to bacterium colony different shape (surface, edge etc.),
Size counts respectively, homomorphosis, it is of the same size be denoted as one kind, a little bacterium in oese picking single bacterium colony, by " oblique line
Method " is seeded to MRS solid mediums with the single bacterium colony isolated and purified；Single bacterium on bacterium toothpick picking MRS solid mediums
Fall a little bacterium, be seeded to MRS fluid nutrient mediums, be placed in 37 DEG C of incubators, Anaerobic culturel 24h-48h, filters out new bacterial strain.
Embodiment 2 (identification and preservation of lactobacillus gasseri RD-0046 bacterial strains)
1st, cultural character, dyeing microscopic examination and morphological feature：The bacterium colony obtained after culture such as attached drawing 1, bacterium colony is circular, edge
Neatly, surface is smooth, more flat, translucent；The bacterium pure culture smear is taken to carry out Gram's staining, as a result as attached drawing 2, leather are blue
Albert'stain Albert is positive, thin rod shape, holds blunt circle, is arranged in chain or filament shape, the results showed that：Separated bacterial strain preliminary judgement is breast
Bacillus.
2nd, 16SrDNA gene orders are identified：DNA extractions are carried out with bacterial genomes DNA extraction kit, and use primer
To 27F (5 '-AGAGTTTGATCMTGGCTCAG-3 '), 1492R (5 '-TACGGYTACCTTGTTACGACTT-3 '), carries out PCR
Amplification, takes PCR product to carry out gel electrophoresis, determines 16SrDNA genetic fragments, single clearly PCR is about being obtained at 1500bp
Product band, is shown in attached drawing 3.PCR product is purified and determined dna sequence, using Sanger PCR sequencing PCRs, sequencing primer pair
For 27F/1492R, instrument ABI3730XL is sequenced, sequencing result is compared with GenBank databases, it is homologous with lactobacillus gasseri
Property similarity be more than 99%.The category kind finally identified is lactobacillus gasseri.The variable region sequences of its 16SrDNA are shown in sequence table SEQ
ID NO:1。
3rd, physiological and biochemical property：Pass through aesculin hydrolysis experiment, methyl red test (MR experiments), voges-Proskauer test
(VP experiments), indole experiment, triple sugar iron test, kirschner disaccharide iron tests, urease test, phenylalanine deaminase experiment,
Amino acid decarboxylase enzyme test, gelatin liquefaction test, sodium malonate experiment, citrate experiment (citrate experiment), nitrate
Reduction test, litmus milk experiment, bacterium dynamic test measure bacterial strain biochemical reactions, acquired results are as shown in table 1：
The physio-biochemical characteristics experimental result of 1 lactobacillus gasseri RD0046 of table
| Physiology and biochemistry project | As a result |
| Aesculin hydrolysis experiment | + |
| Methyl red test (MR experiments) | + |
| Voges-Proskauer test (VP experiments) | - |
| Indole is tested | - |
| Triple sugar iron test | - |
| Kirschner disaccharide iron tests | - |
| Urease test | - |
| Phenylalanine deaminase is tested | - |
| Amino acid decarboxylase enzyme test | - |
| Gelatin liquefaction test | - |
| Sodium malonate is tested | - |
| Citrate tests (citrate experiment) | - |
| Nitrate reduction test | - |
| Litmus milk is tested | Solidification production acid |
| Bacterium dynamic test | - |
+：Represent positive；-：Represent negative
Biochemical identification, mirror are carried out to bacterial strain using the API 50CHL lactobacillus identification systems of French Mei Liai companies production
It is as shown in table 2 to determine result statistics
2 lactobacillus gasseri RD-0046API 50CH test bar reaction results of table
+：Represent positive；-：Represent negative；d：Represent weakly positive
Thus biochemical collection of illustrative plates judges that bacterial strain biochemical characteristic meets the biochemical characteristic of lactobacillus gasseri.
3rd, the preservation of bacterial strain
Lactobacillus gasseri (Lactobacillus gasseri) RD-0046 of the present invention is cloudy from Chinese healthy women of reproductive age
Screening is got in road secretion, has been preserved in on May 10th, 2017 common in China Committee for Culture Collection of Microorganisms
Microorganism center (abbreviation CGMCC), depositary institution address are：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences
Institute of microbiology, preservation registration number are CGMCC No.14109, and the Classification And Nomenclature of the bacterial strain is lactobacillus gasseri
(Lactobacillus gasseri)。
Embodiment 3 (lactobacillus gasseri RD-0046 metabolites measure)
1. lactic acid content measures in lactobacillus gasseri RD-0046 metabolites：Using high-efficient liquid phase technique (0.005M sulfuric acid water
(0.28ml sulfuric acid (98%) -1000ml water, pH are about that 2.1) lactic acid of this strain culturing 24-48h zymotic fluids is examined to solution
Survey, as a result indicate that lactic acid (L/D lactic acid summation) is about 35mg/mL, far above commercially available Lactobacillus delbrueckii (about 15mg/mL) and often
The lactic acid content of lactobacillus gasseri is advised, lactate detection collection of illustrative plates of the invention is referring to attached drawing 4
2. content of hydrogen peroxide measures in lactobacillus gasseri RD-0046 metabolites：By the peroxide of Mcgroarty etc.
Enzyme process carries out hydrogen peroxide semiquantitative determination, and the lactobacillus gasseri RD-0046 of separated identification is inoculated in H2O2Identification of M RS-
TMB tablets, after 37 DEG C of Anaerobic culturel 24h, take out tablet, expose thalline in atmosphere.Produce H2O2Lactobacillus bacterium colony will be changed into blue
Color, without producing H2O2Bacterium colony is non-discolouring, the H according to Coloring Time to generation2O2Sxemiquantitative is carried out, the result is shown in Fig. 5,5min in figure
When bacterium colony have obvious blue and show, a large amount of bluenesss are obvious during 10min occurs, it is clear that this bacterium is easy to produce hydrogen peroxide, production
Hydrogen peroxide is very capable, these results suggest that this lactobacillus gasseri RD-0046 can produce lactic acid and hydrogen peroxide, helps to tie up
Hold microecology in vaginas balance.
Embodiment 4 (antibiotic sensitivity test)
According to the requirement of antibiotic susceptibility test in the 3rd Tiny ecosystem viable bacteria product introduction of version pharmacopeia in 2010, use
AGP test paper disk method measures the sensitiveness of strains, investigated for 0 generation and passes on RD-0046 pairs of the lactobacillus gasseri in 30 generations
The sensitiveness of each antibiotic, judges strains sensitiveness rank, the measurement result such as institute of table 3 according to the size of inhibition zone
Show：
The antibiotic sensitivity test result of 3 lactobacillus gasseri RD0046 of table
Inhibition zone is determined as slight sensitive less than 10mm, is medium sensitivity in 10-20mm, is sensitivity more than 20mm.
Experimental data shows this bacterium to bacitracin, fleraxacin slight sensitive, to oxacillin, kanamycins clindamycin
With gentamicin medium sensitivity, to ampicillin, benzyl penicillin, tetracycline, erythromycin, Piperacillin, ceftriaxone, mould through the ages
Element, amoxicillin/clavulanate, azithromycin, Amoxicillin, SM 7338 are sensitive.
Attached drawing 6. wherein is shown in the antibiotic sensitive figure of kanamycins, fleraxacin and amoxicillin/clavulanate
Embodiment 5 (toxicity test)
5 SPF grades of Kunming mouses, lactobacillus gasseri RD-0046 suspended bacterias fresh every mouse peritoneal injection 0.3ml
Liquid (is more than 1 × 109CFU/ mouse).By 2015 editions Chinese Pharmacopoeia requirements, every mouse weight is measured daily, and is observed, recorded
The change such as the front and rear behavior of every mouse injection and physiology.All the weight of animals have increase in the results show 7 days, have no obvious
Poisoning symptom, crawler behavior is without exception, no animal dead, it is believed that the bacterial strain belongs to non-toxic type bacterial strain.
Embodiment 6 (test of lactobacillus gasseri RD-0046 mitotic stabilities)
The present embodiment is from growth characteristics, morphology, Biochemical Characteristics, metabolin component, antibiotic sensitive characteristic, hereditary capacity
And toxotest etc. has carried out this lactobacillus gasseri bacterial strain RD-0046 the study on the stability of passage 30 generations (C30).
1st, lactobacillus gasseri RD-0046 isolate and purify, colony morphological observation, dyeing microscopic examination and biochemical characteristic detection method it is same
The Part I of embodiment 1 and embodiment 2.The result shows that：By passage, colonial morphology is shown in for circle, neat in edge, surface light
Sliding, more flat, translucent, passage is stablized；Gram's staining is rendered as Gram-positive bacillus, and dyeing microscopic examination photo does not have with 0 generation
Change.
2nd, Genetic Analysis：Part II of the method with embodiment 2.Respectively to the 0th generation of lactobacillus gasseri RD-0046
(C0), the 30th generation (C30) bacterial strain carries out 16SrDNA fragment PCR amplifications, and pcr amplification product electrophoretic analysis, purpose band is clear
And it is single, size is about 1500bp, and amplification is correct, and C0, C30 twice PCR amplification are consistent, will measure sequence and uses in NCBI
BLAST instruments be compared with the known array in GenBank databases, be lactobacillus gasseri, homology similarity
100%.
3rd, metabolite measures：Method is each with embodiment 3, Plasma lactate content about 35mg/mL, hydrogen peroxide experiment display
Occurs blueness in 5min for bacterium colony, a large amount of bluenesss are obvious during 10min occurs, it was demonstrated that bacterial strain metabolism produces hydrogen peroxide
Ability and lactic acid producing ability are stablized.
4th, antibiotic sensitivity test：Method is with embodiment 4, using using AGP test paper disk method measure strains
Sensitiveness, judge the strain of this lactobacillus gasseri to bacitracin, fluorine sieve according to the scope of restraining fungi criteria for interpretation of paper disc method
Husky star slight sensitive, to oxacillin, kanamycins clindamycin and gentamicin medium sensitivity, to ampicillin, penicillin
G, tetracycline, erythromycin, Piperacillin, ceftriaxone, vancomycin, amoxicillin/clavulanate, azithromycin, A Moxi
Woods, SM 7338 are sensitive.
5th, toxicity test：Method with embodiment 5, with mouse peritoneal injection to the C0 of this lactobacillus gasseri RD-0046,
C30, for bacterial strain carried out toxotest, wherein, the concentration ＞ 10 of test9CFU/ mouse.As a result it is：Whole test mices 7 days
Poisoning symptom is inside showed no, weight has increase, no animal dead.According to the above results, press《New drug pharmacology, toxicological study skill
Art requires supplementary notes》, which belongs to non-toxic type bacterial strain.
Comprehensive, the present embodiment repeatedly passes on lactobacillus gasseri RD-0046 with MRS medium cultures, from morphology, biochemistry
Learn, metabolin feature and hereditary capacity, susceptibility characteristic, toxicity test etc. have inquired into the numerous shadow planted to lactobacillus gasseri of passage
Ring.The result shows that：With MRS subcultures within 30 generations its morphology, biochemical, hereditary capacity, metabolin and susceptibility characteristic
It is consistent with initial separation bacterial strain.
Embodiment 7 (lactobacillus gasseri RD-0046 bacterial strains pharmacodynamic experiment)
First, lactobacillus gasseri RD-0046 bacterial strains antibacterial experiment in vitro
(1) lactobacillus gasseri RD-0046 and Lactobacillus delbrueckii suppress the experiment of gardnerella vaginalis in vitro：Connect according to 3%
Kind concentration prepares the MRS agar bacteria cakes of lactobacillus gasseri RD-0046, and the Anaerobic culturel 24h at 37 DEG C, commercially available De Shi is prepared with method
Lactobacillus bacteria cake；100 μ L of gardnerella vaginalis are taken, is inoculated in 10mL BHI solid mediums, is put into lactobacillus gasseri RD-
0046 and Lactobacillus delbrueckii bacteria cake, 37 DEG C of Anaerobic culturel 48h；Obvious inhibition zone occurs around lactobacillus.The result is shown in Fig. 7, its
Middle left figure is lactobacillus gasseri RD-0046 inhibition zone effects, and vernier caliper measurement antibacterial circle diameter is 30.7mm, right figure De Shi
Lactobacillus inhibition zone effect, bacteriostatic diameter 24.6mm, conclusion are lactobacillus gasseri RD-0046 to the antibacterial of gardnerella vaginalis
Effect is better than Lactobacillus delbrueckii.
(2) lactobacillus gasseri RD-0046 and Lactobacillus delbrueckii suppress the experiment of aerobic pathogenic bacteria in vitro：According to 3% inoculation
Concentration prepares the MRS agar bacteria cakes of lactobacillus gasseri RD-0046, and the Anaerobic culturel 24h at 37 DEG C, commercially available De Shi breasts are prepared with method
Bacillus bacteria cake；By staphylococcus aureus, escherichia coli, pseudomonas aeruginosa and salmonella are seeded in pancreas junket soya peptone
Liquid (TSB) agar medium, is put into lactobacillus gasseri RD-0046 and Lactobacillus delbrueckii bacteria cake.18-24h is cultivated in 33 DEG C, is seen
Inhibition zone is examined, vernier caliper measurement RD-0046 antibacterial circle diameters are about 235mm, and right figure is Lactobacillus delbrueckii inhibition zone effect, suppression
The a diameter of 15.3mm of bacterium, conclusion are false single to staphylococcus aureus, escherichia coli, verdigris for lactobacillus gasseri RD-0046
The fungistatic effect of born of the same parents bacterium and salmonella is better than Lactobacillus delbrueckii, and representative diagram is shown in attached drawing 8-9
2nd, adhesive forces are tested：According to the lactobacillus number being attached on vaginal epithelial cell monolayer, determine not
With the Adhesion property of lactobacillus.Method is as follows：Take human vagina epithelial cell Vk2/E6E7 and human cervical cancer epithelial cell
Hela, cell is inoculated in 12 orifice plates with 500,000 per the density in holes, when 48 is small after VK2/E6E7 form monolayer；
Commercially available lactobacillus (Lactobacillus delbrueckii) and lactobacillus gasseri RD-0046 are separately added into the CFU of varying number per hole, adhesion 4 is small
When, gently vibrated on shaking table in adhesion process, each group is respectively equipped with two parallel laboratory tests；After adhesion, 1ml is used
0.05%tritonX-100 cell lysis, is made suspension bacteria liquid, and dilution, takes 100ul bacterium solutions to be equably inoculated in MRS fine jades respectively
On fat culture medium flat plate；After when Anaerobic culturel 48 is small, clone's number of each tablet is counted.
The results show：The 4h adherence rates of lactobacillus gasseri RD-0046 are respectively 42.4% and 48.2%, commercially available similar De Shi
Lactobacillus strain 4h adherence rates are respectively 11.8% and 17.3%, and the adhesive force of lactobacillus gasseri RD-0046 is higher than commercially available similar
Lactobacillus Lactobacillus delbrueckii bacterial strain.
3rd, rabbit vagina field planting experiment
1st, experimental method
4 healthy animals of selection carry out stratified random packet by weight, are divided into 2 groups, positive controls animal 2, tests
Group 2：
Field planting is prepared with lactobacillus gasseri RD-0046：The freeze-drying bacterium powder of lactobacillus gasseri RD-0046 is weighed, it is fixed to make
Plant amount is 106.Control group uses listing product (Lactobacillus delbrueckii capsule), each to add MRS fluid nutrient mediums under aseptic technique
0.5mL, is uniformly mixed, and vagina is implanted into after all being drawn with vaginal administration device.
It is colonized modeling and sampling：After the monkey menstruation of normal menstrual cycle can be observed, continuous 7 days implantation modeling bacterium.
The vagina of animal is once observed weekly, checks the color of vaginal fluid, character and secretory volume and measure vaginal secretion
Thing pH, takes 2 aseptic cotton carrier samplings, wherein a cotton swab is used for vaginal fluid cleannes microscopy, another cotton swab is used for bacterium
Cluster analysis.
The separation and purifying culture of vagina bacterium：By the vaginal fluid of collection, vibrated in 2mL D-Hanks buffer solutions,
Gradient dilution is done with phosphate buffer, is respectively coated on MRS agar plates and at 37 DEG C, under anaerobic condition cultivate 24~
48h.The information such as record colonial morphology, MRS agar plates of ruling again are with the bacterium colony that is purified and carry out biochemical and molecule and reflect
It is fixed.
Molecular biology method identifies (16SrDNA gene sequencings)：The separated bacterial strain being purified into is carried out
Sequence amplification, sequencing and the analysis of 16SrDNA, gel extraction method pair is used by the pcr amplification product for being accredited as 16SrDNA fragments
Purpose fragment is sequenced after purification.The 16SrDNA gene orders measured are used into the BLAST instruments and GenBank in NCBI
Known array in database is compared, and is greater than or equal to 99% when comparing homology, is then accredited as same species.
2nd, experimental result and analysis
Vagina mucosa and secretion overview：Observed once weekly after implantation, it turns out that all experimental animal vaginas
Mucosal secretions do not find obvious exception.
Vaginal fluid pH value measures：After lactobacillus gasseri RD-0046 implantation, most of test group of animals vaginal secretion
Thing pH value is significantly lower than listing product control group, and relative to being decreased obviously before implantation, pH value measurement result is as shown in table 4.
4 lactobacillus gasseri RD-0046 of table influences result of the test to experimental animal vaginal fluid pH
Vaginal fluid cleannes：Compared before and after implantation, control group vagina miscellaneous bacteria quantity substantially increases, and cleannes reduce；
And experimental group lactobacillus gasseri RD-0046 implantation after, vaginal fluid cleannes are substantially better than control group, it is seen that quantity is not
Deng Gram-positive bacterium vaginae, cleannes are apparently higher than control group.Vaginal fluid cleannes judge the result such as institute of table 5
Show.
5 lactobacillus gasseri RD-0046 of table influences result of the test to experimental animal vaginal fluid cleannes
I~III is clean-up performance, and III represents that cleanliness factor is minimum.
Experimental group secretion is carried out amplification to isolate and purify, is analyzed by vaginal flora, from the whole 3 animal the moon of experimental group
Found in road secretion for examination lactobacillus gasseri RD-0046, the information for examination lactobacillus gasseri RD-0046 isolated is as schemed
Shown in 10.It is seen that found in animal subject vaginal fluid for examination lactobacillus gasseri RD-0046, therefore can be with
Find out that lactobacillus gasseri RD-0046 can effectively be colonized in Chinese machin intravaginal.
Embodiment 8 (lactobacillus gasseri RD-0046 freezes preservation and freeze-dried powder stability)
In order to detect survival rates of the lactobacillus gasseri RD-0046 in the case of fermenting and is lyophilized, by lactobacillus gasseri RD-
0046 grows in the modified MRS culture medium of pH 6.5, uses the ferment tank of 100 liters of scales.In plateau harvested earlier
Thalline, its viable count reach about 3 × 109CFU/ml.Thalline is collected by centrifuging, after being washed with phosphate buffer, with jelly
Dry protective agent (including skimmed milk power and sucrose etc.) mixing.Then, mixture is placed in freeze drier and be freeze-dried.Sample
Freezed at -40 DEG C about 2 it is small when, be then dried in vacuo at -20 DEG C 20-30 it is small when, then when 30 DEG C of dry 3-5 are small.Dry powder
Dispensed in the aluminium foil bag for being put into drier, and be stored in 4 DEG C and room temperature (25 DEG C).Pass through tablet meter in the 0th, 1,2,3,6, December
Number measure viable count.
Initial every gram of dry powder of lactobacillus gasseri RD-0046 contains up to 75,000,000,000 viable bacterias (7.5 × 1010Cfu/g), in 2-8
There is optimal storage stability at DEG C.After being stored 6 months at 2-8 DEG C, retain the 77.3% of initial viable count, be shown in Table 6.
6 lactobacillus gasseri RD-0046 freeze-dried powder microbial inoculum stability test results of table
The embodiment of the present invention is described in detail above, but the content is only presently preferred embodiments of the present invention,
It is not intended to limit the invention.All all any modification, equivalent and improvement done in the application range of the present invention etc., should all
Within protection scope of the present invention.
Sequence table
<110>Zhao Xiangjiang
<120>A kind of Lactobacillus rhamnosus and its application for being used to prepare vagina antibacterial medicines
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1506
<212> DNA
<213>Lactobacillus rhamnosus (Lactobacillus rhamnosus)
<400> 1
aatttttgtc caccttagac ggctcgctcc ctaaaagggt tacgccaccg gcttcgggtg 60
ttacaaactc tcatggtgtg acgggcggtg tgtacaaggc ccgggaacgt attcaccgcg 120
gcgtgctgat ccgcgattac tagcgattcc gacttcgtgt aggcgagttg cagcctacag 180
tccgaactga gaatggcttt aagagattag cttgacctcg cggtctcgca actcgttgta 240
ccatccattg tagcacgtgt gtagcccagg tcataagggg catgatgatt tgacgtcatc 300
cccaccttcc tccggtttgt caccggcagt cttactagag tgcccaacta aatgctggca 360
actagtcata agggttgcgc tcgttgcggg acttaaccca acatctcacg acacgagctg 420
acgacaacca tgcaccacct gtcattttgc ccccgaaggg gaaacctgat ctctcaggtg 480
atcaaaagat gtcaagacct ggtaaggttc ttcgcgttgc ttcgaattaa accacatgct 540
ccaccgcttg tgcgggcccc cgtcaattcc tttgagtttc aaccttgcgg tcgtactccc 600
caggcggaat gcttaatgcg ttagctgcgg cactgaaggg cggaaaccct ccaacaccta 660
gcattcatcg tttacggcat ggactaccag ggtatctaat cctgttcgct acccatgctt 720
tcgagcctca gcgtcagtta cagaccagac agccgccttc gccactggtg ttcttccata 780
tatctacgca tttcaccgct acacatggag ttccactgtc ctcttctgca ctcaagtttc 840
ccagtttccg atgcacttcc tcggttaagc cgagggcttt cacatcagac ttaaaaaacc 900
gcctgcgctc gctttacgcc caataaatcc ggataacgct tgccacctac gtattaccgc 960
ggctgctggc acgtagttag ccgtggcttt ctggttggat accgtcacgc cgacaacagt 1020
tactctgccg accattcttc tccaacaaca gagttttacg acccgaaagc cttcttcact 1080
cacgcggcgt tgctccatca gacttgcgtc cattgtggaa gattccctac tgctgcctcc 1140
cgtaggagtt tgggccgtgt ctcagtccca atgtggccga tcaacctctc agttcggcta 1200
cgtatcattg ccttggtgag ccgttacctc accaactagc taatacgccg cgggtccatc 1260
caaaagcgat agcttacgcc atctttcagc caagaaccat gcggttcttg gatttatgcg 1320
gtattagcat ctgtttccaa atgttatccc ccacttaagg gcaggttacc cacgtgttac 1380
tcacccgtcc gccactcgtt caaaattaaa tcaagatgca agcacctttc aataatcaga 1440
actcgttcga cttgcatgta ttaggcacgc cgccagcgtt catcctgacc agaaaaaaaa 1500
actcat 1506
Claims (9)
1. a kind of lactobacillus gasseri, its deposit number is CGMCC No.14109.
2. a kind of DNA molecular, it is characterised in that the DNA molecular extracts from lactobacillus gasseri described in claim 1, using it as mould
Version is expanded using 16s-rDNA universal primers, amplified production sequence and lactobacillus gasseri base sequence two in GenBank databases
Person's homology is more than 98%.
3. lactobacillus gasseri described in claim 1 is used to prepare the application of vaginal pathogenic antibacterial medicines.
4. application according to claim 3, it is characterised in that the medicine is microbial inoculum, which colonizes in vaginal environment
And survive on vaginal epithelial cell.
5. application according to claim 3, it is characterised in that the medicine is microbial inoculum, which is metabolized in vaginal environment
Produce H2O2。
6. application according to claim 3, it is characterised in that the pathogenic bacteria are Gardnerella vaginalis or atropic ripple bacterium.
7. application according to claim 3, it is characterised in that the pathogenic bacteria are Candida albicans, Staphylococcus aureus
Bacterium, escherichia coli, pseudomonas aeruginosa or salmonella.
8. lactobacillus gasseri described in claim 1 is used to prepare vaginal disease prevention or the application of medicine.
9. application according to claim 8, it is characterised in that the vaginal disease is vulvovaginal candidiasis, drop
Worm property vaginitis, senile vahinitis, non-specific vagina infection or Combination vagina infection.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810026176.2A CN108004187B (en) | 2018-01-11 | 2018-01-11 | Lactobacillus gasseri and application thereof in preparing vagina bacteriostatic drug |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810026176.2A CN108004187B (en) | 2018-01-11 | 2018-01-11 | Lactobacillus gasseri and application thereof in preparing vagina bacteriostatic drug |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108004187A true CN108004187A (en) | 2018-05-08 |
| CN108004187B CN108004187B (en) | 2020-03-10 |
Family
ID=62049985
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810026176.2A Active CN108004187B (en) | 2018-01-11 | 2018-01-11 | Lactobacillus gasseri and application thereof in preparing vagina bacteriostatic drug |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108004187B (en) |
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402002A (en) * | 2018-11-01 | 2019-03-01 | 南京工业大学 | Lactobacillus gasseri and application thereof in preparation of premature delivery prevention medicine |
| CN110016442A (en) * | 2019-02-21 | 2019-07-16 | 河北一然生物科技有限公司 | The application of Lactobacillus rhamnosus and its composite bacterium powder in prevention and treatment vaginitis product |
| CN110540945A (en) * | 2018-05-29 | 2019-12-06 | 广东强基药业有限公司 | lactobacillus jensenii and application thereof in preparation of vaginal antibacterial drugs |
| CN110538201A (en) * | 2018-05-28 | 2019-12-06 | 赵湘江 | Lactobacillus composition and application thereof |
| CN111088178A (en) * | 2018-10-23 | 2020-05-01 | 上海上药信谊药厂有限公司 | Produce lactic acid and H2O2Lactobacillus and application thereof |
| CN111471623A (en) * | 2020-04-21 | 2020-07-31 | 广东龙创基药业有限公司 | Composition of three lactobacilli and application thereof |
| WO2021027740A1 (en) | 2019-08-09 | 2021-02-18 | 四川厌氧生物科技有限责任公司 | Multi-lactobacillus composition and application thereof to vaginal health of females |
| CN112458006A (en) * | 2020-11-10 | 2021-03-09 | 深圳华大生命科学研究院 | Lactobacillus gasseri for the prevention and/or treatment of diseases associated with disturbances of the genital flora |
| CN113862188A (en) * | 2021-10-15 | 2021-12-31 | 广东一元兰欣生物科技有限公司 | Lactobacillus gasseri LS03 and application thereof |
| CN114214256A (en) * | 2022-01-12 | 2022-03-22 | 哈尔滨美华生物技术股份有限公司 | Lactobacillus gasseri for preventing and treating urogenital infection and application thereof |
| CN114250186A (en) * | 2022-01-05 | 2022-03-29 | 江南大学 | Lactobacillus gasseri for relieving bacterial vaginitis and application thereof |
| CN114350560A (en) * | 2022-01-05 | 2022-04-15 | 江南大学 | Lactobacillus paracasei for inhibiting growth and biomembrane of gardnerella vaginalis and producing hydrogen peroxide at high yield |
| CN116948922A (en) * | 2023-09-20 | 2023-10-27 | 杭州微致生物科技有限公司 | Lactobacillus gasseri VB247 and application thereof |
| CN117327628A (en) * | 2023-11-16 | 2024-01-02 | 江苏科荣生物医药有限公司 | Lactobacillus gasseri Mia capable of fermenting Chinese herbal medicine and resisting HPV virus and application thereof |
| CN117535207A (en) * | 2024-01-04 | 2024-02-09 | 四川厌氧生物科技有限责任公司 | Lactobacillus gasseri and application thereof |
| CN118109375A (en) * | 2024-04-30 | 2024-05-31 | 善恩康生物科技（苏州）有限公司 | Lactobacillus gasseri JM6 and application thereof in vaginitis |
| WO2024146601A1 (en) * | 2023-01-06 | 2024-07-11 | 上海上药信谊药厂有限公司 | Lactobacillus gasseri and use thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080299099A1 (en) * | 2007-06-04 | 2008-12-04 | Instytut Biotechnologii Surowic I Szczepionek Biomed S.A. | Strain composition of the lactobacillus genus and the application of strain composition of the lactobacillus genus |
| EP2364712A1 (en) * | 2010-03-11 | 2011-09-14 | Kloarys Developpement | Topical cosmetic or pharmaceutical composition comprising probiotic lactobacillus strains and use of same |
| CN103911309A (en) * | 2013-01-05 | 2014-07-09 | 欣力菲生物医药技术（天津）有限公司 | Lactobacillus gasseri strain and its uses |
| CN103911306A (en) * | 2013-01-05 | 2014-07-09 | 欣力菲生物医药技术（天津）有限公司 | Lactobacillus gasseri strain and uses thereof |
-
2018
- 2018-01-11 CN CN201810026176.2A patent/CN108004187B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080299099A1 (en) * | 2007-06-04 | 2008-12-04 | Instytut Biotechnologii Surowic I Szczepionek Biomed S.A. | Strain composition of the lactobacillus genus and the application of strain composition of the lactobacillus genus |
| EP2364712A1 (en) * | 2010-03-11 | 2011-09-14 | Kloarys Developpement | Topical cosmetic or pharmaceutical composition comprising probiotic lactobacillus strains and use of same |
| CN103911309A (en) * | 2013-01-05 | 2014-07-09 | 欣力菲生物医药技术（天津）有限公司 | Lactobacillus gasseri strain and its uses |
| CN103911306A (en) * | 2013-01-05 | 2014-07-09 | 欣力菲生物医药技术（天津）有限公司 | Lactobacillus gasseri strain and uses thereof |
Non-Patent Citations (1)
| Title |
|---|
| MARK WILKS等: "Identification and H2O2 Production of Vaginal Lactobacilli from Pregnant Women at High Risk of Preterm Birth and Relation with Outcome", 《JOURNAL OF CLINICAL MICROBIOLOGY》 * |
Cited By (23)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110538201A (en) * | 2018-05-28 | 2019-12-06 | 赵湘江 | Lactobacillus composition and application thereof |
| CN110540945A (en) * | 2018-05-29 | 2019-12-06 | 广东强基药业有限公司 | lactobacillus jensenii and application thereof in preparation of vaginal antibacterial drugs |
| CN111088178A (en) * | 2018-10-23 | 2020-05-01 | 上海上药信谊药厂有限公司 | Produce lactic acid and H2O2Lactobacillus and application thereof |
| CN111088178B (en) * | 2018-10-23 | 2022-11-22 | 上海上药信谊药厂有限公司 | Produce lactic acid and H 2 O 2 Lactobacillus and application thereof |
| CN109402002A (en) * | 2018-11-01 | 2019-03-01 | 南京工业大学 | Lactobacillus gasseri and application thereof in preparation of premature delivery prevention medicine |
| CN110016442A (en) * | 2019-02-21 | 2019-07-16 | 河北一然生物科技有限公司 | The application of Lactobacillus rhamnosus and its composite bacterium powder in prevention and treatment vaginitis product |
| WO2021027740A1 (en) | 2019-08-09 | 2021-02-18 | 四川厌氧生物科技有限责任公司 | Multi-lactobacillus composition and application thereof to vaginal health of females |
| CN111471623A (en) * | 2020-04-21 | 2020-07-31 | 广东龙创基药业有限公司 | Composition of three lactobacilli and application thereof |
| WO2022100633A1 (en) * | 2020-11-10 | 2022-05-19 | 深圳华大生命科学研究院 | Lactobacillus gasseri for prevention and/or treatment of reproductive tract flora disorder-related diseases |
| CN112458006A (en) * | 2020-11-10 | 2021-03-09 | 深圳华大生命科学研究院 | Lactobacillus gasseri for the prevention and/or treatment of diseases associated with disturbances of the genital flora |
| CN113862188A (en) * | 2021-10-15 | 2021-12-31 | 广东一元兰欣生物科技有限公司 | Lactobacillus gasseri LS03 and application thereof |
| CN114250186A (en) * | 2022-01-05 | 2022-03-29 | 江南大学 | Lactobacillus gasseri for relieving bacterial vaginitis and application thereof |
| CN114350560A (en) * | 2022-01-05 | 2022-04-15 | 江南大学 | Lactobacillus paracasei for inhibiting growth and biomembrane of gardnerella vaginalis and producing hydrogen peroxide at high yield |
| CN114350560B (en) * | 2022-01-05 | 2023-07-04 | 江南大学 | Lactobacillus paracasei capable of inhibiting growth and biomembrane of gardnerella vaginalis and producing hydrogen peroxide |
| CN114214256B (en) * | 2022-01-12 | 2022-06-28 | 哈尔滨美华生物技术股份有限公司 | Lactobacillus gasseri for preventing and treating urogenital infection and application thereof |
| CN114214256A (en) * | 2022-01-12 | 2022-03-22 | 哈尔滨美华生物技术股份有限公司 | Lactobacillus gasseri for preventing and treating urogenital infection and application thereof |
| WO2024146601A1 (en) * | 2023-01-06 | 2024-07-11 | 上海上药信谊药厂有限公司 | Lactobacillus gasseri and use thereof |
| CN116948922A (en) * | 2023-09-20 | 2023-10-27 | 杭州微致生物科技有限公司 | Lactobacillus gasseri VB247 and application thereof |
| CN116948922B (en) * | 2023-09-20 | 2024-03-01 | 杭州微致生物科技有限公司 | Lactobacillus gasseri VB247 and application thereof |
| CN117327628A (en) * | 2023-11-16 | 2024-01-02 | 江苏科荣生物医药有限公司 | Lactobacillus gasseri Mia capable of fermenting Chinese herbal medicine and resisting HPV virus and application thereof |
| CN117327628B (en) * | 2023-11-16 | 2024-03-22 | 江苏科荣生物医药有限公司 | Lactobacillus gasseri Mia capable of fermenting Chinese herbal medicine and resisting HPV virus and application thereof |
| CN117535207A (en) * | 2024-01-04 | 2024-02-09 | 四川厌氧生物科技有限责任公司 | Lactobacillus gasseri and application thereof |
| CN118109375A (en) * | 2024-04-30 | 2024-05-31 | 善恩康生物科技（苏州）有限公司 | Lactobacillus gasseri JM6 and application thereof in vaginitis |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108004187B (en) | 2020-03-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108004187A (en) | A kind of lactobacillus gasseri and its application for being used to prepare vagina antibacterial medicines | |
| CN107299065A (en) | Lactobacillus plantarum and application thereof in preparation of vagina bacteriostatic drugs | |
| US9937214B2 (en) | Lactobacillus crispatus and application thereof | |
| CN104630083B (en) | A kind of Lactobacillus crispatus and its application in feminine care products | |
| CN110656060B (en) | Multi-linked lactobacillus composition and application thereof in female vaginal health | |
| CN108004188A (en) | A kind of Lactobacillus rhamnosus and its application for being used to prepare vagina antibacterial medicines | |
| CN107267415B (en) | Lactobacillus reuteri and application thereof in preparing vagina bacteriostatic drug | |
| CN110777087B (en) | Lactobacillus johnsonii and application thereof | |
| CN104178437A (en) | Lactobacillus crispatus and its application in gynecological diseases | |
| WO2021027742A1 (en) | Lactobacillus crispatus and application thereof | |
| CN110540945B (en) | Lactobacillus jensenii and application thereof in preparing vaginal antibacterial drugs | |
| CN117535208B (en) | Lactobacillus crispatus and application thereof in female genital tract health | |
| CN117535207A (en) | Lactobacillus gasseri and application thereof | |
| CN111471623A (en) | Composition of three lactobacilli and application thereof | |
| CN111154677B (en) | Lactobacillus acidophilus and application thereof | |
| CN117511826B (en) | Lactobacillus mucilaginosus and application thereof | |
| CN117535206B (en) | Lactobacillus salivarius and application thereof | |
| CN117535209B (en) | Lactobacillus brevis and application thereof in female genital tract health | |
| CN117363511B (en) | Webster-fused-type medicine capable of improving colpitis diseases and application thereof | |
| CN110538201A (en) | Lactobacillus composition and application thereof | |
| CN118726132A (en) | Canine lactobacillus composite microecological preparation and application thereof | |
| CN118185803A (en) | Lactobacillus reuteri ML23 and application thereof | |
| CN116536177A (en) | Lactobacillus rhamnosus GB99 with bowel relaxing function |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP02 | Change in the address of a patent holder | ||
| CP02 | Change in the address of a patent holder |
Address after: 521000 self built R & D Building 1 / F and 2 / F, plot jn02-15-1, shenchao Road, Jingnan sub Park, Zhongshan (Chaozhou) industrial transfer industrial park, Chaozhou City, Guangdong Province Patentee after: GUANGDONG LONGCHUANGJI PHARMACEUTICAL Co.,Ltd. Patentee after: Zhao Xiangjiang Address before: 521000 Chaozhou Avenue, Chaozhou City, Guangdong Province, North End of Chaozhou Avenue, East Chaozhou Economic Development Zone Management Committee Office Building, Fifth Floor, 535 Rooms Patentee before: GUANGDONG LONGCHUANGJI PHARMACEUTICAL Co.,Ltd. Patentee before: Zhao Xiangjiang |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240319 Address after: 521000 Factory Building on Plot JN02-15-1, Shenchao Road, Jingnan Branch, Zhongshan (Chaozhou) Industrial Transfer Industrial Park, Fengquanhu High tech Zone, Chaozhou City, Guangdong Province Patentee after: GUANGDONG LONGCHUANGJI PHARMACEUTICAL CO.,LTD. Country or region after: Zhong Guo Address before: 521000 self built R & D Building 1 / F and 2 / F, plot jn02-15-1, shenchao Road, Jingnan sub Park, Zhongshan (Chaozhou) industrial transfer industrial park, Chaozhou City, Guangdong Province Patentee before: GUANGDONG LONGCHUANGJI PHARMACEUTICAL CO.,LTD. Country or region before: Zhong Guo Patentee before: Zhao Xiangjiang |