CN110538201A

CN110538201A - Lactobacillus composition and application thereof

Info

Publication number: CN110538201A
Application number: CN201810519343.7A
Authority: CN
Inventors: 赵湘江; 潘立
Original assignee: Individual
Current assignee: Guangdong Longchuangji Pharmaceutical Co ltd
Priority date: 2018-05-28
Filing date: 2018-05-28
Publication date: 2019-12-06

Abstract

The invention discloses a lactobacillus composition which contains lactobacillus rhamnosus strain and lactobacillus gasseri strain and can resist bacterial vaginosis and various vaginal infections. The composition has obvious effect on bacterial vaginosis, is safe and nontoxic, has good stability and can be stored for a long time. Compared with other current commercial preparations, the bacterial strains in the composition can be effectively planted, can synergistically play a role after being implanted into a vagina, can form vaginal microecology, and has more obvious effect compared with a single bacterial strain.

Description

Lactobacillus composition and application thereof

Technical Field

the invention belongs to the technical field of medicines, and particularly relates to a lactobacillus composition and application thereof, in particular to a medicine for treating gynecological diseases.

background

a plurality of microorganisms exist in the vagina of healthy women, and the microorganisms, a host and the environment form a vaginal microecosystem which is mutually restricted, coordinated and dynamically balanced. The vaginal flora of healthy women is mainly composed of lactobacillus, including lactobacillus rhamnosus, lactobacillus jensenii, lactobacillus gasseri, lactobacillus crispatus, lactobacillus vaginalis, etc. Under normal conditions, the lactobacillus can protect the vagina, and when the vaginal microecological lactobacillus disorder can cause pathogenic bacteria to invade to generate vaginitis.

Bacterial Vaginosis (BV) occurs because of imbalance of vaginal flora and reduction of host lactobacillus, which leads to mass propagation of other conditional pathogenic microorganisms such as Gardner bacteria, various anaerobes, Vibrio flexuosus and the like, and BV is usually a mixed infection mainly comprising Gardner bacteria. The use of antibiotics to treat BV temporarily alleviates the symptoms of BV, but also causes the already reduced lactobacilli to be further reduced, aggravates vaginal dysbiosis, and thus allows BV to recur repeatedly. How to control relapse and radically cure bacterial vaginosis is a delicate problem which needs to be solved urgently by gynecologists.

The existing products on the market at present are not the dominant bacterial flora in the vagina of women in China, have poor planting capability, can not maintain stable viable bacterial count and can not completely meet the clinical requirements of gynecology. The vagina of a healthy woman contains a plurality of lactobacilli, and a single strain is difficult to play a good role when being used as a vaginal flora for regulation, and even if the single strain has good acid production capacity, H2O2 production capacity and the capability of adhering to vaginal epithelial cells, the single strain is difficult to re-fix and enter the vaginal flora, and is difficult to play a role for a long time. When a plurality of strains are successfully planted in the vagina, the synergistic effect is an important basis for the continuous effect of the lactobacillus, is also a key factor for the curative effect of the lactobacillus, and can effectively form a vaginal microecosystem.

Disclosure of Invention

The invention aims to solve the technical problem of providing a lactobacillus composition which can effectively and synergistically play a role, wherein the strain of the composition is from a healthy human body, has active and stable biological characteristics and is a combination of lactobacillus rhamnosus and lactobacillus gasseri with strong bacteriostatic ability.

The technical scheme of the invention is as follows:

A lactobacillus composition comprises a lactobacillus rhamnosus strain and a lactobacillus gasseri strain, wherein the lactobacillus rhamnosus (RD-0060) has the preservation number of CGMCC No.14184, and is preserved in the China general microbiological culture Collection center; the preservation number of the Lactobacillus gasseri (RD-0046) is CGMCC No.14109, and the Lactobacillus gasseri is preserved in the China general microbiological culture Collection center of the culture Collection of microorganisms.

Further, the composition can also comprise pharmaceutically acceptable pharmaceutic adjuvants such as lactose, microcrystalline cellulose, starch, pre-crosslinked starch, silicon dioxide, magnesium stearate and the like, and can also not contain pharmaceutic adjuvants.

The lactobacillus composition is used for preparing medicines for preventing and/or treating gynecological diseases.

The application of the lactobacillus composition in preparing a medicament for preventing and/or treating vaginal diseases.

Furthermore, the pathogenic bacteria of the vaginal diseases comprise one or more of Gardnerella, Clostridium, Candida albicans, atrophaera, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa and Salmonella.

the application of the lactobacillus composition in preparing a medicament with the function of adhering vaginal epithelial cells.

The lactobacillus composition is used for preparing a medicament for regulating the balance of vaginal flora.

Two Lactobacillus rhamnosus RD-060 and Lactobacillus gasseri RD-0046 of the invention are screened from vaginal secretions of healthy women of China, and a large number of experiments prove that the Lactobacillus rhamnosus RD-060 and the Lactobacillus gasseri RD-0046 of the invention have stronger acid-producing, H2O 2-producing and vaginal epithelial cell adhesion capabilities. The medicine for preventing and/or treating the gynecological diseases, in particular to the medicine for preventing and/or treating vaginal pathogenic bacteria, the medicine for preventing and/or treating vaginal diseases, the medicine for regulating vaginal flora balance and the medicine with the function of adhering vaginal epithelial cells can play a good medicinal effect due to the characteristics of the medicine.

The beneficial effect that adopts above-mentioned technical scheme to produce lies in: (1) the Lactobacillus rhamnosus RD-060 and Lactobacillus gasseri RD-0046 strains of the invention can independently and synergistically resist bacterial vaginosis and various vaginal infections, including Candida albicans vaginitis, gonorrhea, viral vaginitis, urinary tract infection and the like. (2) The strain is directly collected from a healthy human body, has active and stable biological characteristics, does not need domestication and rejuvenation processes, and can be directly prepared by a preparation process. (3) The strain has the effects of inhibiting Gardnerella, clostridium, Candida albicans, atrophaera, staphylococcus aureus, Escherichia coli, pseudomonas aeruginosa, salmonella and the like, has obvious advantages compared with the current commercially available preparation, has strong field planting capability when implanted, and can effectively play a role. (4) Can effectively form vaginal microecology, which is obviously different from the planting effect of single strain.

Preservation information

The Lactobacillus rhamnosus RD-060 is screened from vaginal secretions of women of healthy child-bearing age in China, wherein the Lactobacillus rhamnosus RD-0060 is deposited in China general microbiological culture collection center of China microbiological culture collection management Committee, the preservation place is Beijing in China, the preservation number is CGMCC No.14184, and the preservation date is 2017, 05 and 24 days; wherein the Lactobacillus gasseri RD-0046 preservation unit is the common microorganism center of China Committee for culture Collection of microorganisms, the preservation place is Beijing, China, the preservation number is CGMCC No.14109, and the preservation date is 2017, 05 and 10 days.

Drawings

FIG. 1: composition bacteriostatic cake pattern-staphylococcus aureus;

FIG. 2: the composition is bacteriostatic, namely clostridium;

FIG. 3: a result graph of extraction of the genomic DNA of Lactobacillus gasseri RD-0046;

FIG. 4: a PCR product amplification result chart of the Lactobacillus gasseri RD-0046;

FIG. 5: a genomic DNA extraction result chart of lactobacillus rhamnosus RD-0060;

FIG. 6: and (3) a PCR product amplification result chart of the lactobacillus rhamnosus RD-0060.

Detailed Description

The test reagent consumables used in the following examples are all conventional biochemical reagents unless otherwise specified; the experimental methods are conventional methods unless otherwise specified; in the following examples,% represents mass%.

The bacterial culture media used in the following examples can be prepared as follows:

1. broth solid Medium (MRS) preparation:

(1) preparing agar powder into a solution, and adding 1.5g/100ml deionized water;

(2) Adding 17.91g/100ml agar solution of MRS culture medium, and mixing;

(3) Placing into a sterilizing pot, and sterilizing for 20 minutes by 1.0MPa steam;

(4) Pouring the culture medium into a culture dish after the temperature of the culture medium is reduced to room temperature, wherein the culture medium is about 10 ml/piece or 20 ml/piece according to the size of the culture dish;

(5) operating in a clean bench. Cooling to become agar, marking the name of the culture medium and the preparation date, and placing in a refrigerator at 4 ℃ for later use.

2. Broth liquid Medium (MRS) preparation:

(1) Adding the MRS culture medium into deionized water in a ratio of 17.9l g/100 ml;

(2) Placing into a sterilizing pot, and sterilizing for 20 minutes by 1.0MPa steam;

(3) Taking out the sterilized pan when the pressure of the sterilized pan is not high, and subpackaging the sterilized pan into EP tubes with each 1.0 ml. Marking the name of the culture medium and the preparation date, and placing the culture medium in a refrigerator at 4 ℃ for later use.

example 1 preparation of lyophilized powder of Lactobacillus rhamnosus RD-060

Lactobacillus rhamnosus RD-060 was grown in broth liquid Medium (MRS) at pH 6.5, fermented using a 100 liter scale fermenter, and cultured anaerobically. The thallus is collected in the early stage of the plateau stage, and the viable count reaches more than about 1.0 multiplied by 109 CFU/ml. The cells were collected by centrifugation, washed with phosphate buffer and mixed with lyoprotectants (including skimmed milk powder and sucrose). The mixture was then freeze-dried in a freeze-dryer. The samples were frozen at-40 ℃ for about 2 hours, then vacuum dried at-20 ℃ for 20-30 hours, and then dried at 30 ℃ for 3-5 hours. And subpackaging the dry powder in an aluminum foil bag filled with a drying agent, and refrigerating for storage. Counting viable bacteria of the lyophilized powder, wherein the viable bacteria count should be above 1.0 × 1010 CFU/g.

example 2 preparation of lyophilized powder of Lactobacillus gasseri RD-0046

Lactobacillus gasseri RD-0046 was grown in broth liquid culture (MRS) pH 6.5, fermented using a 100 liter scale fermenter, and cultured anaerobically. The thallus is collected in the early stage of the plateau stage, and the viable count reaches more than about 1.0 multiplied by 109 CFU/ml. The cells were collected by centrifugation, washed with phosphate buffer and mixed with lyoprotectants (including skimmed milk powder and sucrose). The mixture was then freeze-dried in a freeze-dryer. The samples were frozen at-40 ℃ for about 2 hours, then vacuum dried at-20 ℃ for 20-30 hours, and then dried at 30 ℃ for 3-5 hours. And subpackaging the dry powder in an aluminum foil bag filled with a drying agent, and refrigerating for storage. Counting viable bacteria of the lyophilized powder, wherein the viable bacteria count should be above 1.0 × 1010 CFU/g.

example 3 preparation of the composition

Respectively mixing 100g of lactobacillus rhamnosus RD-060 lyophilized powder, 100g of lactobacillus gasseri RD-0046 powder, 2.3kg of microcrystalline cellulose and 10g of magnesium stearate, filling into No. 2 hard capsules, filling 220 mg-250 mg of each capsule, preparing into lactobacillus composition capsules, and refrigerating and storing. Counting confirms that the viable count of single bacteria is more than 1.0 × 108 CFU/g.

example 4 preparation of the composition

respectively mixing 10g of lactobacillus rhamnosus RD-060 freeze-dried powder, 100g of lactobacillus gasseri RD-0046 powder, 2.4kg of microcrystalline cellulose and 10g of silicon dioxide, filling into No. 2 hard capsules, filling 220 mg-250 mg of each capsule, preparing into lactobacillus composition capsules, and refrigerating and storing. Counting confirms that the viable count of the lactobacillus rhamnosus RD-060 is more than 1.0 multiplied by 107CFU/g, and the viable count of the lactobacillus gasseri RD-0046 is more than 1.0 multiplied by 108 CFU/g.

example 5 preparation of the composition

100g of lactobacillus rhamnosus RD-060 freeze-dried powder, 10g of lactobacillus gasseri RD-0046 powder, 2.4kg of microcrystalline cellulose and 10g of magnesium stearate are respectively mixed and filled into No. 2 hard capsules, each capsule is filled with 220 mg-250 mg, and the lactobacillus composition capsules are prepared and refrigerated for storage. Counting confirms that the viable count of the lactobacillus rhamnosus RD-060 is more than 1.0 multiplied by 108CFU/g, and the viable count of the lactobacillus gasseri RD-0046 is more than 1.0 multiplied by 107 CFU/g.

example 6 preparation of the composition

100g of lactobacillus rhamnosus RD-060 lyophilized powder, 100g of lactobacillus gasseri RD-0046 powder, 2.3kg of lactose and 10g of magnesium stearate are respectively mixed and filled into No. 2 hard capsules, each capsule is filled with 220 mg-250 mg to prepare lactobacillus composition capsules, and the capsules are refrigerated and stored. Counting confirms that the viable count of a single bacterium is more than 1.0 multiplied by 108 CFU/g.

Example 7 preparation of the composition

Respectively mixing 10g of lactobacillus rhamnosus RD-060 lyophilized powder, 100g of lactobacillus gasseri RD-0046 powder, 2.4kg of lactose and 10g of magnesium stearate, filling into No. 2 hard capsules, filling 220 mg-250 mg of each capsule, preparing into lactobacillus composition capsules, and refrigerating and storing. Counting confirms that the viable count of the lactobacillus rhamnosus RD-060 is more than 1.0 multiplied by 107CFU/g, and the viable count of the lactobacillus gasseri RD-0046 is more than 1.0 multiplied by 108 CFU/g.

Example 8 preparation of the composition

Example 9 toxicity examination of the composition

5 mice of Kunming species of SPF grade were vaginally administered 0.3ml of Lactobacillus composition suspension (more than 1X 109 CFU/mouse) per mouse. The body weight of each mouse was measured daily according to the requirements of chinese pharmacopoeia 2015, and the behavior, physiology, etc. of each mouse before and after injection were observed and recorded. The results show that the weight average of all animals is increased within 7 days, no obvious toxic symptoms are seen, the activities are not abnormal, no animals die, and the lactobacillus composition is considered to be nontoxic.

Example 10 bacteriostatic ability test of composition against pathogenic bacteria

The vagina inflammation is often related to pathogenic bacteria, such as Gardnerella, Clostridium, Candida albicans, atrophaera, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella and the like, so that the composition, the single bacteria and the marketed product (Dijunsheng) are respectively investigated with the Staphylococcus aureus and the Clostridium in an in vitro bacteriostasis test.

Firstly, investigation of bacteriostatic ability of staphylococcus aureus

Lactobacillus rhamnosus RD-060, Lactobacillus gasseri RD-0046, compositions (Lactobacillus rhamnosus RD-060 and Lactobacillus gasseri RD-0046), marketed products (dijunsheng) were separately cultured in broth solid Medium (MRS) for 48h, and 8mm solid cake culture was formed with a round sampler.

The golden staphylococcus liquid is coated in a mannitol sodium chloride agar plate, then the fungus cake is placed in a solid culture medium containing the golden staphylococcus, and is cultured for 24 hours at 37 ℃, the size of a bacteriostasis zone is observed for 24 hours, and the results are as follows:

——	Composition comprising a metal oxide and a metal oxide	Lactobacillus rhamnosus	Lactobacillus gasseri	Marketable product (Dingjunsheng)
					Size of the fungus ring	15.4mm	14.3mm	13.2mm	10.7mm

The bacteriostatic bacterial cake of the composition is shown in figure 1 in the attached figure of the specification.

the bacteriostatic ability of staphylococcus aureus which is an aerobic pathogen is respectively the composition (lactobacillus rhamnosus RD-060 and lactobacillus gasseri RD-0046) > lactobacillus rhamnosus RD-060> lactobacillus gasseri RD-0046> marketed product (Dijunsheng).

Investigation of bacteriostatic ability of clostridium

The clostridium liquid is coated in a Columbia agar culture medium plate, then the fungus cake is placed in the Columbia agar culture medium plate containing the clostridium, and the fungus cake is anaerobically cultured for 24 hours at 37 ℃, the size of a bacteriostasis zone is observed for 24 hours, and the result is as follows:

——	Composition comprising a metal oxide and a metal oxide	Lactobacillus rhamnosus	Lactobacillus gasseri	marketable product (Dingjunsheng)
					Size of the fungus ring	35.3mm	29.8mm	30.2mm	26.4mm

Wherein the fungus cake pattern of the composition is shown in figure 2 of the attached figure of the specification.

The bacteriostatic ability against clostridia was composition (lactobacillus rhamnosus RD-060 and lactobacillus gasseri RD-0046) > lactobacillus rhamnosus RD-060> lactobacillus gasseri RD-0046> marketed (dijunsheng), respectively.

EXAMPLE 11 vaginal colonization experiments on compositions

4 healthy animals were selected and divided into 2 groups by weight, 2 animals in the control group (animals numbered XX-01 and XX-02) and 2 animals in the experimental group (animals numbered XX-03 and XX-04). Wherein the experimental animal is female cynomolgus monkey provided by Zhongke laboratory animals Co., Ltd, West mountain, Suzhou. The test method is as follows:

After monkey menstruation observed in normal menstrual cycle, metronidazole suppositories (200 mg/suppository) were vaginally administered for 5 consecutive days, and then the capsules prepared in example 3 were implanted with bacteria for 5 consecutive days. The vagina of the animal is observed once a week, the color, the character and the secretion amount of the vaginal secretion are checked, the pH value of the vaginal secretion is measured, 2 aseptic polyester swabs are sampled, one swab is used for microscopic examination of the cleanness of the vaginal secretion, and the other swab is used for flora analysis.

Separation and purification culture of vaginal bacteria: the collected vaginal secretion is shaken in 2mL of D-Hanks buffer solution, and is subjected to gradient dilution by phosphate buffer solution, and the obtained product is respectively coated on MRS agar plates and cultured for 24-48 h at 37 ℃ under an anaerobic condition. Single colony morphology and the like are recorded, and the MRS agar plate is restreaked to obtain purified colonies and subjected to biochemical and molecular identification.

Molecular biological method identification (16SrDNA gene sequence analysis): and (3) carrying out 16SrDNA sequence amplification, sequencing and analysis on the separated and purified strain, and carrying out sequencing after a PCR amplification product identified as a 16SrDNA fragment is purified on a target fragment by using a gel cutting recovery method. The determined 16SrDNA gene sequence is aligned with known sequences in GenBank databases using the BLAST tool in NCBI, and the same species is identified when the alignment homology is greater than or equal to 99%.

Experimental results and analysis of vaginal mucosa and secretions general observations: after the implantation, the vaginal mucosa secretion of all the test animals is observed once a week, and the result shows that no obvious abnormality is found.

(1) Measuring the pH value of vaginal secretion: after the capsule administration, the pH value of vaginal secretion of most experimental animals is obviously lower than that of a control group (Dingjunsheng) on the market, and is obviously reduced relative to that before implantation, and the measurement results of the pH value are shown in the following table:

(2) Cleanliness of vaginal secretions: compared with the control group before and after implantation, the vaginal cleanliness of the control group is slightly reduced; after the test group is implanted with the drug, the cleanliness of vaginal secretion is obviously better than that of a control group, gram-positive vaginal bacilli with different quantities can be seen, and the cleanliness is obviously higher than that of the control group. The results of the determination of the cleanliness of vaginal secretions are shown in the following table:

I to III represent the cleanliness, and III represents the lowest cleanliness.

The secretion of the experimental group is amplified, separated and purified, and after vaginal flora analysis, Lactobacillus rhamnosus RD-0060 and Lactobacillus grignard (Lactobacillus gasseri) RD-0046 are found in all the vaginal secretions of 2 animals in the experimental group, the information of the separated Lactobacillus is shown as Lactobacillus rhamnosus RD-0060 and Lactobacillus grignard (Lactobacillus gasseri) RD-0046, and the sequencing information refers to nucleotide and amino acid sequence lists, so that the Lactobacillus rhamnosus RD-0060 and Lactobacillus grignard (Lactobacillus gasseri) RD-0046 can be effectively planted in the vagina of the cynomolgus monkey.

The comparison shows that the cleanliness and the pH value of the composition after field planting are obviously superior to those of the marketed Dingjunsheng, the separation shows that the composition can be effectively field-planted, and the compound double bacteria can also generate a synergistic effect in the vagina to form an ecological flora.

electrophorogram of PCR amplification products of 16SrDNA fragments separated by two strains in fixed planting:

The extraction result of the genomic DNA of Lactobacillus gasseri RD-0046 is shown in the attached figure 3 of the specification;

The result of PCR amplification of Lactobacillus gasseri RD-0046 is shown in FIG. 4 of the accompanying drawings.

The extraction result of the Lactobacillus rhamnosus RD-0060 genome DNA is shown in figure 5 attached to the instruction book;

the result of PCR amplification of Lactobacillus rhamnosus RD-0060 is shown in FIG. 6.

Example 12 cell adhesion of compositions

The composition can be used for treating bacterial vaginitis. When the medicine is used in vagina, the medicine will adhere to vaginal epithelial cell to produce effect.

Culturing a single strain RD-0046\ RD-0060 in an MRS culture medium, respectively incubating the composition preparation with VK2/E6E7 human vaginal epithelial cells cultured in vitro for 2h and 4h after culturing the composition preparation in the MRS culture medium, cleaning non-adhered bacteria after culturing, then cracking the cells by triton x-100 to prepare a suspension bacterial liquid, inoculating the suspension bacterial liquid on an MRS agar culture plate after gradient dilution, counting the colony number of each plate after anaerobic culture for 48h, calculating the adhesion rate, and dividing the adhered bacteria by the initial bacteria to obtain the adhesion rate.

The data show that, in combination, there is mutual promotion of adhesion to the cells, which has a positive effect on the synergistic effect.

Sequence listing

<110> Zhao Xiang Jiang

<120> a lactobacillus composition and use thereof

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<170> SIPOSequenceListing 1.0

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<213> Lactobacillus gasseri (Lactobacillus gasseri)

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gcgtgctgat ccgcgattac tagcgattcc agcttcgtgt aggcgagttg cagcctacag 180

ttcgaactga gaacggcttt cagagatccg cttaccttcg caggttcgct tctcgttgta 240

ccgtccattg tagcacgtgt gtagcccagg tcataagggg catgatgact tgacgtcatc 300

cccaccttcc tccggtttgt caccggcagt ctcattagag tgcccaactt aatgatggca 360

actaatgaca agggttgcgc tcgttgcggg acttaaccca acatctcacg acacgagctg 420

acgacagcca tgcaccacct gtctcagcgt ccccgaaggg aacacctaat ctcttaggtt 480

tgcactggat gtcaagacct ggtaaggttc ttcgcgttgc ttcgaattaa accacatgct 540

ccaccgcttg tgcgggcccc cgtcaattcc tttgagtttc aaccttgcgg tcgtactccc 600

caggcggagt gcttaatgcg ttagctgcag cactgagagg cggaaacctc ccaacactta 660

gcactcatcg tttacggcat ggactaccag ggtatctaat cctgttcgct acccatgctt 720

tcgagcctca gcgtcagttg cagaccagag agccgccttc gccactggtg ttcttccata 780

tatctacgca ttccaccgct acacatggag ttccactctc ctcttctgca ctcaagttca 840

acagtttctg atgcaattct ccggttgagc cgaaggcttt cacatcagac ttattgaacc 900

gcctgcactc gctttacgcc caataaatcc ggacaacgct tgccacctac gtattaccgc 960

ggctgctggc acgtagttag ccgtgacttt ctaagtaatt accgtcaaat aaaggccagt 1020

tactacctct atctttcttc actaccaaca gagctttacg agccgaaacc cttcttcact 1080

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gcgtgctgat ccgcgattac tagcgattcc gacttcgtgt aggcgagttg cagcctacag 180

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ccatccattg tagcacgtgt gtagcccagg tcataagggg catgatgatt tgacgtcatc 300

cccaccttcc tccggtttgt caccggcagt cttactagag tgcccaacta aatgctggca 360

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atcaaaagat gtcaagacct ggtaaggttc ttcgcgttgc ttcgaattaa accacatgct 540

ccaccgcttg tgcgggcccc cgtcaattcc tttgagtttc aaccttgcgg tcgtactccc 600

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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

1. A lactobacillus composition comprises a lactobacillus rhamnosus strain and a lactobacillus gasseri strain, and is characterized in that the lactobacillus rhamnosus (RD-0060) has the preservation number of CGMCC No.14184, and is preserved in the China general microbiological culture Collection center; the preservation number of the Lactobacillus gasseri (RD-0046) is CGMCC No.14109, and the Lactobacillus gasseri is preserved in the China general microbiological culture Collection center.

2. A pharmaceutical composition comprising the lactobacillus composition of claim 1 and a pharmaceutically acceptable excipient.

3. The pharmaceutical composition of claim 2, wherein the pharmaceutically acceptable excipient comprises lactose, microcrystalline cellulose, starch, pre-crosslinked starch, silicon dioxide, magnesium stearate.

4. use of a lactobacillus composition according to any of claims 1 to 3 for the preparation of a medicament for the prevention and/or treatment of gynaecological disorders.

5. The lactobacillus composition for use according to claim 4, wherein the gynecological disease medicament comprises vaginal diseases.

6. Use according to claim 5, characterized in that the pathogenic bacteria of said vaginal diseases comprise Denax bacteria, Candida albicans, Atomobacter, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella.

7. Use of a lactobacillus composition according to any of claims 1 to 3 for the preparation of a medicament with the effect of modulating vaginal flora balance.