Vaginal microbiome composition

Abstract

The present invention provides compositions and methods for treating patients. [Solution] The exemplary composition may include a vaginal microbiome composition. The vaginal microbiome composition may include a mixture of bacteria. The bacterial mixture may include Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, and Lactobacillus rhamnosus.

Description

[Technical Field]

[0001] This disclosure relates to compositions and methods for treating patients. [Background technology]

[0002] A wide variety of compositions and methods can be used to treat diseases and / or conditions, such as the reproductive system, the digestive tract, and the liver. , developed to treat diseases and / or conditions of the immune system, etc. Known compositions and Each method has its own specific advantages and disadvantages. There is an ongoing need to provide alternative compositions and methods. [Overview of the project] [Means for solving the problem]

[0003] This disclosure provides alternative designs, materials, and manufacturing methods for compositions and methods used to treat patients. , and use is provided. A vaginal microbiome composition is disclosed. The vaginal microbiome composition is lac Lactobacillus crispatus, Lactobacillus crispatus Lactobacillus iners, Lactobacillus... Lactobacillus gasseri, Lactobacillus genseni Lactobacillus jensenii, Lactobacillus reuteri (L Actobacillus reuteri, Lactobacillus acidophilus (La Lactobacillus acidophilus, Lactobacillus vaginalis (L Actobacillus vaginalis, Lactobacillus rhamnosus (L Actobacillus rhamnosus, Lactobacillus johnsonii (L Lactobacillus johnsonii), Lactobacillus helveticus (Lactobacillus helveticus), Lactobacillus plantarum (Lactobacillus plantarum), Lactobacillus fermentum (Lactobacillus fermentum), Lactobacillus salivarius (Lactobacillus salivarius), and bacteria including Lactobacillus delbrueckii (Lactobacillus delbrueckii) (including a mixture).

[0004] Instead of or in addition to any of the above embodiments, the bacterial mixture is 25 - 75% of Lactobacillus crispatus (including).

[0005] Instead of or in addition to any of the above embodiments, the bacterial mixture is 30 - 60% of Lactobacillus crispatus (including).

[0006] Instead of or in addition to any of the above embodiments, the bacterial mixture is 40% of Lacto bacillus crispatus (Lactobacillus crispatus) including .

[0007] Instead of or in addition to any of the above embodiments, the bacterial mixture is 10 - 40% of Lactobacillus iners (Lactobacillus iners) including.

[0008] Instead of or in addition to any of the above embodiments, the bacterial mixture is 15 - 30% It contains Lactobacillus iners.

[0009] In place of or in addition to any of the above embodiments, the bacterial mixture is 20% lact This includes Lactobacillus iners.

[0010] In place of or in addition to any of the above embodiments, the bacterial mixture is 1-10% Contains Lactobacillus gasseri.

[0011] In place of or in addition to any of the above embodiments, the bacterial mixture may be 2-8% Includes Lactobacillus gasseri.

[0012] In place of or in addition to any of the above embodiments, the bacterial mixture contains 5% lact Includes Bacillus gasseri.

[0013] In place of or in addition to any of the above embodiments, the bacterial mixture is 1-10% Includes Lactobacillus jensenii nothing.

[0014] In place of or in addition to any of the above embodiments, the bacterial mixture may be 2-8% Includes Lactobacillus jensenii .

[0015] In place of or in addition to any of the above embodiments, the bacterial mixture contains 5% lact Includes Bacillus jensenii.

[0016] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% It contains Lactobacillus reuteri.

[0017] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% It contains Lactobacillus reuteri.

[0018] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Includes Bacillus reuteri (Lactobacillus reuteri).

[0019] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus acidophilus (including us)

[0020] In place of or in addition to any of the above embodiments, the bacterial mixture is approximately 0.5 to 4 % Lactobacillus acidophilus (includes lus)

[0021] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Bacillus acidophilus include.

[0022] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus vaginalis include.

[0023] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% Lactobacillus vaginalis include.

[0024] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Includes Bacillus vaginalis (Lactobacillus vaginalis).

[0025] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus rhamnosus Includes.

[0026] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% Lactobacillus rhamnosus Includes.

[0027] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Includes Bacillus rhamnosus.

[0028] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus johnsonii Includes.

[0029] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% Lactobacillus johnsonii Includes.

[0030] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Includes Bacillus johnsonii.

[0031] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus helvetic (including us)

[0032] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% Lactobacillus helvetic (including us)

[0033] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Bacillus helveticus include.

[0034] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus plantarum Includes.

[0035] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% Lactobacillus plantarum Includes.

[0036] In place of or in addition to any of the above embodiments, the bacterial mixture contains about 1% lactate. Includes Tobacillus plantarum .

[0037] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus fermentum ) includes.

[0038] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% Lactobacillus fermentum ) includes.

[0039] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Includes Bacillus fermentum (Lactobacillus fermentum) .

[0040] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus salivarius ) includes.

[0041] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% Lactobacillus salivarius ) includes.

[0042] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Includes Bacillus salvarius (Lactobacillus salivarius) .

[0043] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.1-5% Lactobacillus delbruec Includes kii.

[0044] In place of or in addition to any of the above embodiments, the bacterial mixture is 0.5-4% Lactobacillus delbruec Includes kii.

[0045] In place of or in addition to any of the above embodiments, the bacterial mixture contains 1% lact Bacillus delbrueckii Includes.

[0046] In place of or in addition to any of the above embodiments, the bacterial mixture contains one or more bacteria. Includes the freeze-dried strain shown above.

[0047] In place of or in addition to any of the above embodiments, the bacterial mixture is contained in the capsule. It will be placed.

[0048] In place of, or in addition to, any of the above embodiments, the bacterial mixture is placed in the suppository. It will be done.

[0049] In place of or in addition to any of the above embodiments, the bacterial mixture is a soluble shell It will be placed inside.

[0050] A method for treating infectious diseases is disclosed. This method is one of the embodiments described above. This includes administering a vaginal microbiome composition to a patient with an infection.

[0051] In place of or in addition to any of the above embodiments, the infection may be bacterial vaginosis, candidiasis, etc. Dust mite infection, human papillomavirus infection, urinary tract infection, sexually transmitted infection, and gynecological cancer cysts Contains one or more of the following:

[0052] A method for treating a patient is disclosed. This method is one of the embodiments described above. This includes administering a vaginal microbiome composition to a patient.

[0053] In place of, or in addition to, any of the above embodiments, the vaginal microbiome composition is administered to the patient. The benefits include preventing premature birth, preventing miscarriage, treating infertility, and interstitial This includes treating one or more of the following: treating cystic cystitis and treating polycystic ovary syndrome.

[0054] A vaginal microbiome composition is disclosed. The vaginal microbiome composition is provided in a suppository and placed inside the suppository. It contains a mixture of freeze-dried bacteria, and the mixture of freeze-dried bacteria includes Lactobacillus crispata Lactobacillus crispatus, Lactobacillus inas ( Lactobacillus iners, Lactobacillus gasseri (Lactoba Lactobacillus gasseri, Lactobacillus gensenii Lactobacillus jensenii, Lactobacillus reuteri Lactobacillus acidophilus (Lactobacillus reuteri) acidophilus, Lactobacillus vaginalis Lactobacillus rhamnosus (Lactobacillus vaginalis) Lactobacillus johnsonii (Lactobacillus s rhamnosus) Lactobacillus helveticus (Lactobacil) Lactobacillus helveticus, Lactobacillus plantarum (Lactobaci Lactobacillus plantarum, Lactobacillus fermentum Lactobacillus fermentum, Lactobacillus salvarius Illus salivarius), and Lactobacillus delbrückii (Lac Includes *Tobacillus delbrueckii*.

[0055] A vaginal microbiome composition bank is disclosed. The vaginal microbiome composition bank is arranged within A first container containing a first population of one or more microorganisms, wherein the first population is a first A first container collected from a na, and a second population of one or more microorganisms placed inside it. A second container containing a second population collected from a second donor, Includes an index system containing donor data from a first donor and a second donor. nothing.

[0056] A vaginal microbiome composition bank is disclosed. The vaginal microbiome composition bank is arranged within A first container containing a first population of one or more microorganisms, wherein the first population is a first A first container collected from a na, and a second population of one or more microorganisms placed inside it. A second container containing a second population collected from a second donor, Includes.

[0057] In place of or in addition to any of the above embodiments, the vaginal microbiome composition bank is: Includes an index system containing donor data from the first and second donors.

[0058] A vaginal microbiome composition bank is disclosed. The vaginal microbiome composition bank is arranged within A first container containing a first population of one or more freeze-dried microorganisms, wherein the first population A first container, collected from a first donor, and one or more freeze-dried microorganisms placed inside it. A second container containing a second population of organisms, the second population being collected from a second donor. , a second container, and an index containing donor data from the first and second donors. The system includes

[0059] A vaginal microbiome composition bank is disclosed. The vaginal microbiome composition bank is arranged within A first container containing a first population of one or more freeze-dried microorganisms, wherein the first population A first container, collected from a first donor, and one or more freeze-dried microorganisms placed inside it. A second container containing a second population of organisms, the second population being collected from a second donor. , including a second container.

[0060] In place of or in addition to any of the above embodiments, the vaginal microbiome composition bank is: Includes an index system containing donor data from the first and second donors.

[0061] The above summaries of some embodiments do not describe each disclosed embodiment or all implementations of this disclosure. It is not intended to clarify. The detailed description that follows will further elaborate on these embodiments. To illustrate with a physical example. [Modes for carrying out the invention]

[0062] Terms defined below may differ from those defined elsewhere in the claims or this specification. Unless otherwise specified, these definitions shall apply.

[0063] All figures, whether explicitly stated or not, are approximate. Therefore, it is assumed to be modified. The term "approximately" generally refers to the value stated by a person skilled in the art. It refers to a range of numbers that are considered equivalent (for example, having the same function or result). In many cases, The term "approximately" may include a number rounded to its maximum number of significant figures.

[0064] When specifying a numerical range using endpoints, it includes all numbers within that range (for example, 1 to 5 includes 1, (Including 1.5, 2, 2.75, 3, 3.80, 4 and 5).

[0065] As used herein and in the appended claims, the singular forms "a", "an", and "The" refers to multiple objects unless the context explicitly indicates otherwise. When used in this document and the attached claims, the term "or" is not used unless the context indicates otherwise. Unless explicitly stated otherwise, it is used in a sense that includes "and / or".

[0066] In the specification, phrases such as "a certain embodiment," "several embodiments," and "other embodiments" The reference indicates that the described embodiments may include one or more specific features, structures, and / or characteristics. It should be noted that this indicates that not all embodiments are specific. This does not necessarily mean that it includes the features, structure, and / or characteristics of [the specified]. When the features, structure, and / or characteristics are described in relation to one embodiment, Such features, structures, and / or characteristics shall not be explicitly stated unless the opposite is explicitly stated. Whether or not they are described, they may be used in conjunction with other embodiments. It should be understood.

[0067] The human microbiome (or human microflora) is found on the surface and deep layers of human skin. , normally present in saliva and oral mucosa, conjunctiva, and in the gastrointestinal tract, genitourinary tract, and / or vaginal canal. It is a microbial aggregate, including microorganisms related to reproductive health, such as the placental microbiome. The human microbiome consists of bacteria, but may also include fungal phages, viruses, and archaea. Some of these organisms perform tasks that are useful to the human host, but humans The functions of most organisms that make up the microbiome are still under investigation. Under normal conditions, These microorganisms do not cause disease in the human host; instead, they are involved in maintaining health.

[0068] The vaginal microbiome can play a role in many different health conditions. Examples include bacterial vaginosis, candidiasis, human papillomavirus infection, and urinary tract infections. Infections, and / or sexually transmitted infections, are examples of other conditions. Human cancers (e.g., cervical cancer), premature birth, miscarriage, infertility, interstitial cystitis, polycystic ovary syndrome These include restoring, normalizing, and / or other aspects of the vaginal microbiome. Shifting by law may help mitigate these and other conditions. Compositions and methods for treating patients are disclosed. At least some of the compositions are The method includes a vaginal microbiome composition. At least some of the methods treat bacterial vaginosis and candidiasis. Treating mites infections, treating human papillomavirus infections, treating urinary tract infections, sexual To treat infections, gynecological cancers (e.g., cervical cancer), prevent premature birth, and prevent miscarriage. Methods to treat infertility, interstitial cystitis, and polycystic ovary syndrome. Includes, etc.

[0069] In healthy women of reproductive age, bacteria of the Lactobacillus genus dominate the vaginal microbiome. There is a tendency for four species to be consistently identified across women, and these are lactose. Bacillus crispatus, Lactobacillus crispatus Bacillus iners, Lactobacillus mo Japanese parsley (Lactobacillus gasseri) and Lactobacillus gencea This is Lactobacillus jensenii. These species are white, Listed in order of frequency of domination among Black, Asian, and Hispanic women. It is generally believed that the dominance of Lactobacillus species in the vaginal microbiome is associated with health status. The dominance of a single genus or species creates a microbiome with low diversity. The increase is associated with a higher risk of ill health and infection. For example, the genus Sn. (eathia), Megasphaera, Atopobium Peptoniphilus (Peptoniphyllus pobium), Dialistar The genus Dialister, the genus Prevotella, and / or guardian. Bacteria from the genus Gardnerella tend to correlate with an unhealthy vaginal microbiome. This is possible because they have been identified in women using other indicators of vaginal ill health. This is due to a high Nugent score and vaginal pH. However, these generalizations are, This can vary among different groups.

[0070] The vaginal microbiome can change between pre-pubescent, pre-menopausal, and post-menopausal women. For example, Women before puberty have a relatively low relative abundance of Lactobacillus and a relatively high microbial diversity. It may have (as well as a relatively thin vaginal epithelium, a relatively thin mucosal layer, and a relatively low amount of glyco (May possess the gene). Premenopausal women have relatively high Lactobacillus abundance and relatively low It may have microbial diversity (and also a relatively thick vaginal epithelium, a relatively thick mucosal layer, and a relatively high (They may have estrogen levels and relatively low amounts of glycogen.) Postmenopausal women are moderately The relative abundance of Lactobacillus is at a certain level, and the microbial diversity is relatively low or lower. (and also a relatively thin vaginal epithelium, a relatively thin mucosal layer, and a relatively moderate amount of estrogen) Bell (and may have a relatively moderate amount of glycogen). Vaginal infections (e.g., bacterial vaginal infections) Postmenopausal women exhibiting symptoms of inflammation tend to have lower Lactobacillus levels and relatively high microbial activity. It can have diversity. In summary, it includes the relative abundance of Lactobacillus, or other factors. Compositions that tend to increase by law, and compositions that help reduce microbial diversity. It may be useful.

[0071] In at least some examples, the compositions disclosed herein (e.g., vaginal microbiome) The product utilizes the vaginal microbiome collected from vaginal microbiome donors. However, the details Other sources of vaginal microbiota, including bacterial cultures, should be considered. Potential donors should be identified. This allows for many screening processes. In some cases, latent Potential donors may be screened using a health history questionnaire. An example questionnaire is: , is it the same as the one the Red Cross uses for screening potential blood donors? Or similar. In some of these and other examples, potential donors have a general sense of Infections and other conditions can be screened for. Such screenings include These may include blood tests, stool tests, urine tests, and vaginal swab tests. Such tests are H IV. Hepatitis (Type A, Type B, and / or Type C), Syphilis, Clostridium difficile C. difficile, bacterial pathogens, eggs and parasites, and / or other related species This may include testing for the presence of certain substances. These are just examples. The donor's health is Blood tests are performed, stool samples are analyzed, urine samples are analyzed, vaginal discharge is analyzed, and health history is reviewed regularly. It can be monitored by updating regularly. Vaginal swabs are Gram-stained and Nug To assess vaginal health through the ENT scoring method, white blood cell count, and detection of yeast presence Urine and / or vaginal swabs may be collected to test for sexually transmitted infections.

[0072] After it is determined that the potential donor is healthy enough to provide samples, the donor is given a sample. A kit (for example, a self-collection kit) is provided. The collection kit includes a donor instruction sheet. , vaginal DNA / RNA collection kits for sequence analysis (e.g., DNAgeneotek(trademark)) The OMNIgene vaginal sample collection kit (ORM-130) is commercially available, and the sample collection and Transport kit (for example, eSwa, commercially available from Copan Diagnostics) b 480C Copan Liquid Amies Elution Swab C (May include collection and transport systems). Donor You may be asked to sign an informed consent form. Each donor You may be asked to collect two samples. The first sample will be used for sequence analysis. It is possible. For example, after washing hands, using a swab from a vaginal DNA / RNA collection kit, A vaginal sample is collected by inserting a device several inches into the vagina and swabbing the vaginal wall for approximately 20 seconds. This can be done. Then, the swab can be inserted into a tube containing the stabilizing buffer, and the tube / The sample can be further analyzed. The second sample is used to culture vaginal microorganisms. This can be done. For example, a vaginal sample can be collected using a swab from a sample collection and transport kit. This can be done. Next, the swab is placed in a liquid amies solution (for example, this is aerobic, anaerobic) Insert into a tube containing (which can store bacteria that are difficult to culture for up to 48 hours) This can be done. During collection, the donor places the sample tube and material inside the biohazard bag. Next, the samples are left at the designated facilities.

[0073] To process a sample for sequencing (e.g., shotgun sequencing), The first protease (e.g., QIAGEN protease) is used with sterile PCR-certified water. By rehydrating the sample, an 80 mg / mL solution can be produced (this is possible if the sample is 10 or more times (This may include inverting the sample upwards.) Cool the sample (e.g., a vaginal DNA / RNA collection kit sample). Remove from storage and vortex for 30 seconds. Shake the sampling tube three times to collect the sample. The solution containing can be brought to the bottom of the tube. 5 microliters of rehydrated The protease can be added to the collection tube containing the vaginal swab sample. The sample can be collected up to 10 times. It can be inverted upwards. Then the sample is placed in a 50°C incubator for 2 hours in ink. It can be vortexed. Then the sample can be vortexed for 30 seconds. Next Then, shake the sampling tube three times to bring the solution containing the sample to the bottom of the tube. The swab can be removed from the tube (this is done by pressing the swab against one side of the tube and removing the swab). (This may include recovering the sample absorbed by the system.) Two 500 microliter samples It can be divided into 2 mL vials for cryopreservation, and the sample can be used for sequence analysis (this This is carried out at suitable processing / analysis facilities such as Divergent (trademark) in St. Paul, MN. It can be stored at -80°C until it is ready to use.

[0074] To process the sample for culture, the sample (e.g., sample collection and transport kit sample) is subjected to anaerobic conditions. It can be placed in the airlock of the chamber and introduced into the chamber. While the tube is still inside, vortex it vigorously for 5 seconds to release the sample from the tip of the swab. This can be done. This is not diluted, or 10 0 It is a diluted solution. Next, take a swab. It can be extracted. The sample is 100 microliters in a 900 microliter sterile solution. By transferring it to a tube containing physiological saline (0.9%), it can undergo a 10-fold serial dilution. The sample may be vortexed vigorously for 5 seconds and / or mixed by inversion. It is also acceptable. This is 10 -1 It is a dilution. 100 microliters -1 Diluent This is then transferred to a new tube containing 900 microliters of sterile physiological saline (0.9%). Yes, it is possible. The sample may be vortexed vigorously for 5 seconds and / or mixed by inversion. It is also acceptable. This is 10 -2 It is a dilution. The sample is 10 -6 Until diluted with the diluent This process can be repeated.

[0075] There are three methods: smear culture plates, CDC anaerobic bacteria 5% sheep blood agar plates, De Man, Rogosa and Sharpe (MRS) agar plates, and 50% M On an RS agar plate, place the following dilution tubes: 10 -3 , 10 -4 , 10 -5 It can be prepared from Yes, it's possible. Simply put, transfer 100 μL from a suitable tube directly onto the surface of the agar plate, and then... Use a bacterial L-shaped spreader and plate rotor to spread the bacteria over the entire surface of the plate. This can be done. Incubate the plate under anaerobic conditions at 35°C for 24 to 120 hours. This allows you to check the plate daily for the appearance of new colonies. Once the plates begin to dry, wrap them with Parafilm. You can then observe the colonies. If they arrive, the number and morphology of the colonies can be recorded (this is plate capture) (This may include taking photographs to aid in documented verification of colony morphology). Further special Individual colonies can be selected for sexualization and streaked onto a new plate. If necessary, for example, Bifidobacterium selective agar (BSA), Enterococcus Cell agar (ECA), cetrimide agar (CA), mannitol salt agar (MS) Selective media such as A) may also be used.

[0076] In some cases, Gram staining may be used. If this is done, the specimen will be a slide Gram stain. It can be applied to laths. When staining liquid cultures, use a sterile inoculation loop. The culture can be applied directly to the slide as a smear. Colonies from the plate can be stained. To color the slide, apply a droplet of sterile saline solution to the center of the slide using an inoculation loop. This allows for the collection of small, single colonies, which can then be added to a droplet. If possible, gently mix the droplet and the sample to obtain a thin, uniform smear. It can be done. The droplets can be dried. Pour anhydrous methanol for 1-2 minutes, then tap The slide can be fixed with methanol by rinsing with water. Fixed smear Then, a primary staining agent (e.g., crystal violet) can be poured in for 1 minute. Tap water The primary stain can be removed by gently rinsing with water. Pour in a secondary staining agent / modant (e.g., Gram-iodine) for 1 minute, then rinse with tap water. Rinse gently until the solvent moving from the slide becomes colorless (for example, about 3 You can decolorize the slide (for about 60 seconds), then gently rinse the slide with tap water. This can be done. Then, the counterstain can be poured onto the slide for 1 minute (for example, safran). If the results of the initial counterstaining are unsatisfactory, replace with basic fuchsin. (It is possible to do this.) The slide can be washed with tap water and dried. The smear can be immersed in oil. It can be tested under lenses, and the results / observations can be recorded.

[0077] Other sample tests, including catalase testing, can be performed, and catalase testing involves water peroxide. By adding ingredients, Staphylococcus and Streptococcus This may include distinguishing it from cocci. Another sample test that can be performed is blood One example is the plasma coagulation test, which involves adding plasma to distinguish between Staphylococcus and Streptococcus bacteria. This may include doing so.

[0078] Isolates determined to be Lactobacillus have their antibiotic susceptibility profiles Further characterization may be necessary to determine the outcome. This is from Lactobacillus from plates cultured overnight. The Rus colony is composed of 90% iso-sensitest broth and 10% MRS broth. Suspend the suspension in the broth until the amount equals 1 MacFarland turbidity standard solution. This may include: Next, immerse the sterile swab in the inoculum and press it against the wall of the tube, removing any excess. The liquid can be removed. The swab can be streaked onto the entire surface of the agar. (Rotate at least twice to ensure uniform distribution). After absorbing excess moisture, Inhibitory concentration (MIC) test strips can be used (for example, this test strip has a scale at the top). (This may include applying the test strip with the cord facing outwards.) With the entire length of the antibiotic gradient in complete contact with the agar surface, press the test paper onto the agar surface. It can be attached and rearranged as needed. Invert the plate and heat at 35°C for 20-4 It can be incubated for 8 hours (or longer). Interpret these results. It is possible.

[0079] Isolated strains can be frozen and stored. This involves streaking the culture overnight onto a streaking plate. This may include inoculating well-isolated colonies selected from the population. Many colonies If selected, a 96-well plate format may be used. May be used (for example, Lactobacillus tends to use MRS). Culture It can be incubated overnight at 35°C in an anaerobic chamber incubator. If the culture is not cloudy by the next morning, use an additional incubation time. That's fine too. When the culture appears visually cloudy, add 200-300 microliters. The solution contains an antifreeze agent (for example, 50-60 microliters of 50% glycerol). It can then be transferred to a new 96-well plate. The final glycerol concentration is 10%. Yes, it is possible. The 96-well plate can be sealed and stored in a freezer at -80°C.

[0080] As suggested herein, individual microbial strains (e.g., individual strains or bacterial species) It can be isolated from the swab. This can then be selectively cultured on an agar medium, for example, for the desired strain. Plating and then "purifying" the strain as a single colony isolate. This may include one or more processes of the following: Individual microbial strains are grown / cultured and sequenced. Biochemical tests, colony morphology, cell morphology, microscopic mortality, and different representative antibiotics Species can be identified / characterized by their sensitivity to quality. Microbial strains can vary. In some cases, the microbial strain is Lactobacillus crispatus. Lactobacillus crispatus, Lactobacillus inas (iners), Lactobacillus gasser i) Lactobacillus jensenii ), Lactobacillus reuteri, Lactobacillus Lactobacillus acidophilus Lactobacillus vaginalis, Lactobacillus rhamnosus, Lactobacillus johnsonii Lactobacillus helveticus s), Lactobacillus plantarum m), Lactobacillus fermentum Lactobacillus salivar Lactobacillus delbruckii (ius), Lactobacillus delbruckii Gardnerella vaginalis (rueckii) lis), non-Lactobacillus bacteria, Atopobium vaginae ginae), Prevotella bivia, these Other species in the genus, Megasphaera, Sneatia ia), Dialister genus, Peptoniphyllus genus It may include one or more of the following: iphilus, fungi, or combinations thereof.

[0081] The vaginal microbiome composition contains a single isolated strain or a mixture of two or more isolated strains. It is thought that... An example composition is approximately 20-85% Lactobacillus crispatus ( Lactobacillus crispatus), or approximately 25-75% lactobacillus Lactobacillus crispatus, or about 30 ~60% Lactobacillus crispa Lactobacillus crispatus (tus), or approximately 40% Lactobacillus crispatus It may include crispatus. In some of these, and in other examples, (for example) (In addition to or in lieu of any one or more of the microorganisms listed above), exemplary The composition contains approximately 10-50% Lactobacillus inas. Lactobacillus iners), or approximately 10-40% Lactobacillus iners (Lactobaci Lactobacillus iners), or approximately 15-30% Lactobacillus iners (Lacto Lactobacillus iners, or approximately 20% Lactobacillus iners. It may also include *Obacillus iners*. Some of these, and other examples. So (for example, in addition to or instead of any one or more of the microorganisms listed above) ), an example composition is approximately 1-10% Lactobacillus gasseri (Lactobacil). Lactobacillus gasseri, or approximately 2-8% Lactobacillus gasseri (Lactoba Lactobacillus gasseri, or approximately 5% Lactobacillus gasseri. It may also include *Acillus gasseri*. Some of these, and other examples. So (for example, in addition to or instead of any one or more of the microorganisms listed above) ), an example composition is approximately 1-10% Lactobacillus gensenii (Lactoba cillus jensenii), or about 2-8% Lactobacillus jensenii ( Lactobacillus jensenii, or approximately 5% Lactobacillus jensenii This may include Lactobacillus jensenii. In some cases, and in other cases (for example, one or more of the microorganisms listed above) In addition, or alternatively, exemplary compositions include approximately 0.1-5% Lactobacillus luteum Lactobacillus reuteri, or approximately 0.5-4% lactobacillus Bacillus reuteri, or about 1% It may also include Lactobacillus reuteri. Yes. In some of these cases, and in other cases (for example, any of the microorganisms listed above), In addition to or instead of one or more of the above, an exemplary composition may contain approximately 0.1-5% lact Bacillus acidophilus Or about 0.5-4% Lactobacillus acidophilus Lactobacillus acidophilus, or approximately 1% Lactobacillus acidophilus (Lacto It may also contain Bacillus acidophilus. In some of these, In other examples (for instance, in addition to one or more of the microorganisms listed above, or so (Instead of this), an example composition is approximately 0.1-5% Lactobacillus vaginalis (La Lactobacillus vaginalis, or approximately 0.5-4% Lactobacillus • Vaginalis (Lactobacillus vaginalis), or about 1% lactobacillus Including Lactobacillus vaginalis This is also good. In some of these, and in other examples (for example, the microorganisms listed above) In addition to or instead of one or more of the above, the exemplary compositions contain approximately 0.1-5% of the following: Lactobacillus rhamnosus, also It contains approximately 0.5-4% Lactobacillus rhamnosus. amnosus), or about 1% Lactobacillus rhamnosus (Lactobacil It may include lus rhamnosus). In some of these, and in other examples, (For example, in addition to or instead of any one or more of the microorganisms listed above), An example composition is approximately 0.1-5% Lactobacillus johnsonii. Lactobacillus johnsonii), or approximately 0.5-4% Lactobacillus johnsonii - (Lactobacillus johnsonii), or about 1% Lactobacillus • It may include *Lactobacillus johnsonii*. In some of these cases, and in other cases (for example, any one of the microorganisms listed above) In addition to or instead of the above, exemplary compositions include approximately 0.1-5% lactobacilli Lactobacillus helveticus, or Approximately 0.5-4% Lactobacillus helveticus Lactobacillus elveticus, or approximately 1% Lactobacillus elveticus (Lactobacillus elveticus) It may also include Acillus helveticus. In some of these, also In other examples (for instance, in addition to or in addition to one or more of the microorganisms listed above), Alternatively, an exemplary composition may contain approximately 0.1-5% Lactobacillus plantarum (Lac Lactobacillus tobacillus, or approximately 0.5-4% Lactobacillus Plantarum (Lactobacillus plantarum), or about 1% Lactobacillus Includes Lactobacillus plantarum But that's fine. In some of these cases, and in other cases (for example, the microorganisms listed above) In addition to or in lieu of one or more of the above, the exemplary compositions contain approximately 0.1-5% Lactobacillus fermentum , or approximately 0.5-4% Lactobacillus fermentum Lactobacillus fermentum), or about 1% Lactobacillus fermentum It may also include *Acillus fermentum*. In some of these, and others In this example (for example, in addition to or instead of any one or more of the microorganisms listed above), For example, an exemplary composition contains approximately 0.1-5% Lactobacillus salvarius. Lactobacillus salivarius), or approximately 0.5-4% Lactobacillus. Lactobacillus salivarius, or about 1% of the population Lactobacillus salivarius It may include. In some of these, and in other examples (for example, the microorganisms listed above) In addition to or in place of one or more of the substances, the exemplary composition is approximately 0.1 to 5 % Lactobacillus delbrue Lactobacillus delbrückii, or approximately 0.5-4% Lactobacillus delbrückii (Lactoba Lactobacillus delbrueckii), or about 1% Lactobacillus delbrueckii It may also include Lactobacillus delbrueckii.

[0082] An example composition is approximately 40% Lactobacillus crispatus (Lactobacil Lactobacillus crispatus), approximately 20% Lactobacillus inas (Lactoba cillus iners), approximately 5% Lactobacillus gasseri (Lactobacil) Lactobacillus gasseri), approximately 5% Lactobacillus gensenii Illus jensenii), about 1% Lactobacillus reuteri (Lactoba Lactobacillus reuteri), approximately 1% Lactobacillus acidophilus (Lactobacillus acidophilus) Lactobacillus acidophilus, approximately 1% Lactobacillus vaginalis (Lactobacillus vaginalis), approximately 1% Lactobacillus rum Lactobacillus rhamnosus, approximately 1% lactobacillus Lactobacillus johnsonii, approximately 1% Lactobacillus helveticus Approximately 1% of Lactobacillus plantarum Lactobacillus f(rum), approximately 1% Lactobacillus fermentum Lactobacillus salvarius (Lactobacillus ermentum), approximately 1% (us salivarius), and about 1% Lactobacillus delbruckii (La It may also include *Ctobacillus delbrueckii*.

[0083] An example composition is Lactobacillus crispatus (Lactobacillus c Lactobacillus gasseri Lactobacillus jensenii ii), and Lactobacillus rhamnosus It may contain osus). For example, the composition may contain about 20 - 85% of Lactobacillus crispatus, or about 25 - 75% of Lactobacillus crispatus, or about 30 - 60% of Lactobacillus crispatus, or about 40% of Lactobacillus crispatus. The composition may contain about 1 - 10% of Lactobacillus gasseri, or about 2 - 8% of Lactobacillus gasseri, or about 5% of Lactobacillus gasseri. The composition may contain about 1 - 10% of Lactobacillus jensenii, or about 2 - 8% of Lactobacillus jensenii, or about 5% of Lactobacillus jensenii. The composition may contain about 0. 1 - 5% of Lactobacillus rhamnosus, or about 0.5 - 4% of Lactobacillus rhamnosus, or about 1% of Lactobacillus rhamnosus. In some cases, each of the components may be freeze - dried. The total number of microorganisms in an exemplary composition is about 1×10 ~1× 10 CFU, or about 1×10 CFU. In some cases, each of the components may be freeze - dried. The total number of microorganisms in an exemplary composition is about 1×10 5 ~1× 10 15 CFU, or about 1×106 ~1 × 10 12 CFU, or approximately 1 × 10⁻⁶ 7 ~1 × 10 10 It may also be ordered in CFU quantities. The composition may be a capsule, suppository, soluble shell, and / or can be placed in a suitable delivery vehicle such as other similar vehicles. In some cases, the composition can be administered orally. In other cases, the composition can be administered topically (for example, (By inserting capsules, suppositories, soluble shells, etc., into the vagina.)

[0084] An example composition is Lactobacillus crispatus (Lactobacillus crispatus) having the desired properties. Lactobacillus crispatus, Lactobacillus gasseri Lactobacillus gasseri, Lactobacillus gensenii (us jensenii), and Lactobacillus rhamnosus (Lactobacil It may contain strains of Lus rhamnosus. For example, each strain of the component may be pH-lowering. It possesses properties, produces lactic acid, and has inhibitory activity against other microorganisms with other unique activities. They may be selected to have combinations of these, and / or other similar ones. .

[0085] An example composition is freeze-dried Lactobacillus crispatus. Lactobacillus crispatus), freeze-dried Lactobacillus gasseri (Lact Lactobacillus gasseri, freeze-dried Lactobacillus gensenii (Lactobacillus jensenii), and freeze-dried lactobacillus Contains a mixture of Lactobacillus rhamnosus Each component can be isolated from the donor. For example, the composition may contain approximately 20-85% of the donor. Lactobacillus crispatus, also Approximately 25-75% of Lactobacillus crispatus Lactobacillus crispatus (Lactobacillus ispatus), or approximately 30-60% Lactobacillus crispatus (Lactobacillus crispatus) Lactobacillus crispatus, or approximately 40% Lactobacillus crispatus It may also contain (Lactobacillus crispatus). The composition is about 1 ~10% Lactobacillus gasseri, Or about 2-8% Lactobacillus gasser i) or about 5% Lactobacillus gasseri The composition may contain approximately 1-10% Lactobacillus gensenii (La). Lactobacillus jensenii, or approximately 2-8% Lactobacillus jensenii Lactobacillus jensenii, or approximately 5% lactobacillus Lactobacillus jensenii may also be included. The composition contains approximately 0.1-5% Lactobacillus rhamnosus. Lactobacillus rhamnosus (L) or approximately 0.5-4% Lactobacillus rhamnosus (L) Actobacillus rhamnosus, or approximately 1% Lactobacillus rhamnosus It may also contain Lactobacillus rhamnosus. Each can be derived / isolated from a donor sample. The total number of microorganisms in the exemplary composition is approximately 1. ×10 5 ~1 × 10 15 CFU, or approximately 1 × 10⁻⁶6 ~1 × 10 12 CFU, or approximately 1x1 0 7 ~1 × 10 10 It may also be ordered in CFU quantities. The composition may be in the form of capsules, suppositories, or soluble tablets. It can be placed in a suitable delivery vehicle such as a shell and / or other similar items. In some cases, the composition may be administered orally. In other cases, the composition may be administered topically. To obtain (for example, by inserting a capsule, suppository, soluble shell, etc., into the vagina).

[0086] An example composition is freeze-dried Lactobacillus crispatus. Lactobacillus crispatus), freeze-dried Lactobacillus gasseri (Lact Lactobacillus gasseri, freeze-dried Lactobacillus gensenii (Lactobacillus jensenii), and freeze-dried lactobacillus Contains a mixture of Lactobacillus rhamnosus Each component can be isolated from the donor. For example, the composition may contain approximately 20-85% of the donor. Lactobacillus crispatus, also Approximately 25-75% of Lactobacillus crispatus Lactobacillus crispatus (Lactobacillus ispatus), or approximately 30-60% Lactobacillus crispatus (Lactobacillus crispatus) Lactobacillus crispatus, or approximately 40% Lactobacillus crispatus It may also contain (Lactobacillus crispatus). The composition is about 1 ~10% Lactobacillus gasseri, Or about 2-8% Lactobacillus gasser i) or about 5% Lactobacillus gasseri The composition may contain approximately 1-10% Lactobacillus gensenii (La). Lactobacillus jensenii, or approximately 2-8% Lactobacillus jensenii Lactobacillus jensenii, or approximately 5% lactobacillus Lactobacillus jensenii may also be included. The composition contains approximately 0.1-5% Lactobacillus rhamnosus. Lactobacillus rhamnosus (L) or approximately 0.5-4% Lactobacillus rhamnosus (L) Actobacillus rhamnosus, or approximately 1% Lactobacillus rhamnosus It may also contain Lactobacillus rhamnosus. Each can be derived from a bacterial culture. The total number of microorganisms in the exemplary composition is approximately 1 × 10⁶. 5 ~1 × 10 15 CFU, or approximately 1 × 10⁻⁶ 6 ~1 × 10 12 CFU, or approximately 1 × 10⁻⁶ 7 ~ 1 x 10 10 It may also be ordered in CFU quantities. The composition may be in the form of capsules, suppositories, or soluble sachets. It can be placed in a suitable delivery vehicle such as a ru and / or other similar vehicle. In some cases, the composition may be administered orally. In other cases, the composition may be administered topically. For example, by inserting a capsule, suppository, or soluble shell into the vagina.

[0087] An example composition is freeze-dried Lactobacillus crispatus. Lactobacillus crispatus), freeze-dried Lactobacillus gasseri (Lact Lactobacillus gasseri, freeze-dried Lactobacillus gensenii (Lactobacillus jensenii), and freeze-dried lactobacillus Contains a mixture of Lactobacillus rhamnosus Each component can be isolated from the donor. For example, the composition may contain approximately 20-85% of the donor. Lactobacillus crispatus, also Approximately 25-75% of Lactobacillus crispatus Lactobacillus crispatus (Lactobacillus ispatus), or approximately 30-60% Lactobacillus crispatus (Lactobacillus crispatus) Lactobacillus crispatus, or approximately 40% Lactobacillus crispatus It may also contain (Lactobacillus crispatus). The composition is about 1 ~10% Lactobacillus gasseri, Or about 2-8% Lactobacillus gasser i) or about 5% Lactobacillus gasseri The composition may contain approximately 1-10% Lactobacillus gensenii (La). Lactobacillus jensenii, or approximately 2-8% Lactobacillus jensenii Lactobacillus jensenii, or approximately 5% lactobacillus Lactobacillus jensenii may also be included. The composition contains approximately 0.1-5% Lactobacillus rhamnosus. Lactobacillus rhamnosus (L) or approximately 0.5-4% Lactobacillus rhamnosus (L) Actobacillus rhamnosus, or approximately 1% Lactobacillus rhamnosus It may also contain Lactobacillus rhamnosus. One or more of these may be derived / isolated from the donor sample, and one or more components may be cultured in a bacterial culture medium. They may be derived from nutrients. The total number of microorganisms in the exemplary composition is about 1 × 10⁶ 5 ~1×1 0 15 CFU, or approximately 1 × 10⁻⁶ 6 ~1 × 10 12 CFU, or approximately 1 × 10⁻⁶ 7 ~1 × 10 1 0 It may also be ordered in CFU quantities. The composition may be in the form of capsules, suppositories, soluble shells, and / or Alternatively, it can be placed in a suitable delivery vehicle such as another similar vehicle. In some cases, the composition can be administered orally. In other cases, the composition can be administered topically (e.g., (By inserting capsules, suppositories, soluble shells, etc., into the vagina.)

[0088] An example composition is Lactobacillus crispatus (Lactobacillus crispatus) having the desired properties. Lactobacillus crispatus, Lactobacillus gasseri Lactobacillus gasseri, Lactobacillus gensenii (us jensenii), and Lactobacillus rhamnosus (Lactobacil It may contain strains of Lus rhamnosus. For example, each strain of the component may be pH-lowering. It possesses properties, produces lactic acid, and has inhibitory activity against other microorganisms with other unique activities. They may be selected to have combinations of these, and / or other similar ones. .

[0089] As shown above, the total number of microorganisms in the example composition is approximately 1 × 10⁻⁶. 5 ~1 × 1015 CFU, or about 1×10 6 ~1×10 12 CFU, or about 1×10 7 ~1×10 10 CF U may be of the order. This may correspond to the total number of microorganisms in a single capsule, suppository, soluble shell, and / or other similar ones. Alternatively, this may correspond to the total number of microorganisms in a suitable dosage form of the composition (e.g., this may include one or more capsules, suppositories, soluble shells, and / or other similar ones).

[0090] A single capsule, suppository, soluble shell, and / or other similar ones can be described as an encapsulated formulation or dosage that can be administered to a patient. This may include administering a dosage to a patient using a suitable dosing regimen. This may include administering one or more doses to a patient For example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 1 2, 13, 14, 15, 16, 17, 18, 19, 20 times, or more doses can be administered to a patient In some of these and other examples, the encapsulated formulation may be administered to the patient once or more per day. For example, the dosage may be administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 1 0, 11, 12, 13, 14 times, or more per day. In some of these and other examples, the dosage may be administered to the patient once or more per day. For example, the dosage may be administered to the patient 1, 2, 3, 4, 5, 6 times, or more.

[0091] One exemplary dosing regimen may include administering 2 doses twice a day for 2 days to a patient Another exemplary dosing regimen may include administering 4 doses twice a day for 2 days to a patient It can be obtained. Another exemplary dosing regimen may involve administering 4 doses twice a day for 4 days to the patient. It may include. Another exemplary dosing regimen may include once-daily dosing for 8 days for the patient. These are merely examples. In at least some of these examples, each dose may be a frozen dried material containing a mixture of Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii, and Lactobacillus rhamnosus.

[0092] In at least some examples, the vaginal microbiota composition may not contain bacteria from the genus Sneathia, Megasphaera, Atopobium, Peptoniphilus, Dialister, Prevotella, and / or Gardnerella. For example, the vaginal microbiota composition may not contain bacteria from the genus Atopobium, Gardnerella, and Prevotella.

[0093] The vaginal microbiota compositions disclosed herein can be used to treat a number of different health conditions. For example, the vaginal microbiota composition can treat bacterial vaginitis, treat candidiasis infections, treat human papillomavirus infections, treat urinary tract infections, treat sexually transmitted ​​​​​​​​​​​To treat infectious diseases, to treat gynecological cancers (e.g., cervical cancer), to reduce and / or prevent premature births, To reduce and / or prevent miscarriage, treat infertility, treat interstitial cystitis, and polycystic ovary syndrome. It can be used to treat the syndrome, etc.

[0094] The vaginal microbiome composition also includes one or more additional microorganisms (e.g., collected from a donor and single A combination of a wide range of vaginal microbiota, which is combined with or mixed with (separated or otherwise) It is thought to include [unclear]. For the purposes of this disclosure, a broad range of vaginal microbiota is representative of female [unclear]. It can be understood as a collection of microorganisms present in the vagina. A wide range of vaginal microorganisms. The flora is intentionally manipulated to alter the presence or absence of any specific microorganism in the sample. It is not a representative sample, but rather intended to represent the entire population of organisms in the sample. The composition of the vaginal microbiome can vary from person to person. Therefore, there are differences in the broad range of vaginal microbiomes. It is possible. In some cases, the vaginal microbiome composition may consist of Lactobacillus crispatus (La Lactobacillus crispatus, Lactobacillus inas (Lact Lactobacillus iners, Lactobacillus gasseri Lactobacillus gasseri, Lactobacillus gensenii jensenii), Lactobacillus reuteri Lactobacillus acidophilus (Lactobacillus aci dophilus), Lactobacillus vaginalis Lactobacillus ginalis, Lactobacillus rhamnosus amnosus), Lactobacillus jo Lactobacillus helveticus (Lactobacillus hnsonii) Lactobacillus helveticus, Lactobacillus plantarum plantarum), Lactobacillus fermentum Lactobacillus salvarius (Lactobacillus fermentum) Lactobacillus salivarius, Lactobacillus delbrückii It is combined with or mixed with one or more of the following: These are thought to include a broad range of vaginal microbiota to which these are added. At least several In this example, both a broad vaginal microbiome and additional microorganisms were supplied from a vaginal microbiome donor. To be given.

[0095] Bacteria / microorganisms are available in capsules, suppositories, soluble shells, and / or for oral or topical delivery. It may be placed in a suitable container such as other similar items. In some cases, simulate The prepared vaginal fluid may be incorporated into the vaginal microbiome composition. In some of these, and in other examples... Therefore, the vaginal microbiome composition may contain glycogen. Total volume contained in the container / suppository This can be on the order of approximately 100 microliters to 10 milliliters.

[0096] In at least some cases, bacteria / microorganisms in vaginal microbiome compositions are freeze-dried. It may contain bacteria / microorganisms. This is because the bacteria / microorganisms are freeze-dried in a freeze-drying process. May contain bacteria / microorganisms, and / or freeze-dried bacteria / microorganisms, orally delivered or Suitable for local delivery include capsules, suppositories, soluble shells, and / or other similar forms. It can be placed inside a container.

[0097] Treatment of a patient can include administration of the vaginal microbiota composition to the patient. One purpose of the treatment is to normalize the patient's vaginal microbiota by effectively replacing the patient's vaginal microbiota with the vaginal microbiota composition. This can include directly administering the vaginal microbiota composition to the patient's vagina by transplantation, suppository , or another suitable route of administration.

[0098] In at least some examples, a plurality of vaginal microbiota compositions can be stored or banked in a suitable storage device / facility. The storage device can include a temperature control device such as a refrigeration device (e.g., a 4°C refrigeration device), a freezer (e.g., a -20°C freezer), an ultra-low temperature freezer (e.g., a -80°C freezer), or other similar ones. In some examples, an indexing system can also be linked to or otherwise associated with the vaginal microbiota composition bank. The indexing system can include data such as donor data that can be used to match the vaginal microbiota composition from a given donor to a suitable patient. Donor data can include age at collection, reproductive status at collection (pre-menopausal, post-menopausal), race, menstrual period at collection, history of bacterial vaginitis, history of candidiasis infection, history of human papillomavirus, other medical histories (e.g., history of Clostridium difficile (C. difficile) infection, history of cancer diagnosis / treatment, etc.), combinations thereof, and / or other similar ones. A clinician can use the donor data to adjust the treatment regimen for a specific patient. For example, a patient with bacterial vaginitis can be treated with a vaginal microbiota composition derived from a donor who has successfully improved bacterial vaginitis.

[0099] In some cases, donor data may include pregnancy history. For example, some donors may have been pregnant. When there is an actual or recognized increase in fertilization ability, whether sought or otherwise, pregnancy occurs. There are cases where there is a high possibility of vaginal contamination from such donors. The internal microbiome composition addresses the challenges of infertility treatment, aiming to increase or enhance the fertilization capacity of patients. It may be administered to patients who are in a state of ill health. In such cases, the vaginal microbiome composition may be administered in oral capsules. It can be administered as a graft, or the vaginal microbiome composition can be administered as a graft, and / or It may be inserted directly into the patient's vagina by other means.

[0100] U.S. Patent No. 9,675,648 is incorporated herein by reference.

[0101] U.S. Patent No. 9,629,881 is incorporated herein by reference.

[0102] U.S. Patent No. 10,226,431 is incorporated herein by reference.

[0103] The specification of U.S. Patent Application Publication No. 2018 / 0289750 is incorporated herein by reference. It gets included. [Examples]

[0104] This disclosure can be further clarified by referring to the following embodiments, The examples are essentially predictive and serve to illustrate several examples and to illustrate the present disclosure. It does not limit anything to any extent.

[0105] Example 1 A vaginal microbiome composition can be manufactured. The vaginal microbiome composition contains approximately 40% lactobacilli. Lactobacillus crispatus, approximately 2 0% Lactobacillus iners, approximately 5% Lactobacillus gasseri, approximately 5% Lactobacillus jensenii, approximately 1% Lactobacillus reuteri, approximately 1% Lactobacillus acidophilus Lactobacillus vaginalis (Lactobacillus va ginalis), approximately 1% Lactobacillus rhamnosus Lactobacillus johnsonii (Lactobacillus rhamnosus), approximately 1% Illus johnsonii), approximately 1% Lactobacillus helveticus (La Lactobacillus helveticus), approximately 1% Lactobacillus plantae Lactobacillus plantarum, approximately 1% Lactobacillus... Fermentum (Lactobacillus fermentum), approximately 1% lactobacillus Bacillus salivarius, and Approximately 1% Lactobacillus delbruckii Contains a mixture of bacteria, including *Eueckii*.

[0106] A mixture of bacteria can be placed in a suppository. The total volume contained in the suppository (for example, The amount of material containing a mixture of bacteria is approximately 100 microliters to approximately 10 milliliters. It could be made to order.

[0107] This suppository can be administered to patients. For example, the suppository can be used for bacterial vaginosis, candidiasis, and other related conditions. One of the following: infectious diseases, human papillomavirus infection, urinary tract infections, sexually transmitted infections, and gynecological cancers. It can be administered to a patient to treat one or more conditions.

[0108] This suppository can be administered to patients. For example, the suppository can prevent premature birth and miscarriage. To prevent, treat infertility, treat interstitial bladder, treat polycystic ovary syndrome, and / or These combinations can be administered to patients.

[0109] Example 2 - Exemplary Lactobacillus gensenii for use in vaginal microbiome compositions (Lactobacillus jensenii) strain Lactobacillus gensenii, designated as stock number 8001-D01-M006. Donate (e.g., as described herein) the strain of *Obacillus jensenii*. It was isolated from donor specimens deposited (which were donated to be used for this purpose). The donor had a vaginal swallow. Eligibility is obtained by donating blot and urine samples, which are then Gram-stained and Nugent-stained. For scoring (GRAM) Fairview Diagnostics Labor Sent to the atory, next STI: Trichomonas vaginalis nalis) (TVPCR), Neisseria gonorrhoeae ( GCPCR), Chlamydia trachomatis We performed a test for s)(CHPCR).

[0110] During the preparation of the working cell stock of 8001-D01-M006, the cultured cell density was adjusted. Setting this was not targeted, but a single wave from a previously adjusted image coating plate By selecting isolated colonies, overnight culture was initiated. This overnight culture was then cultured the following morning. Dilute with fresh broth in a 1:5 ratio, let grow for at least 4 hours, then glycerinate the culture. Combined with the roll, a 10% glycerol stock was prepared. Generally, in this way... The adjustment of the glycerol stock is approximately 1 x 10 9 Stock with a cell density of CFU / ml It is causing problems.

[0111] OmniGene and ESwab vaginal swab specimens (8001-D01) were subjected to metagenomics. Sequencing (OmniGene), culture, and qPCR identity testing Provided for use as an ESwab. ESwab for culture and qPCR identity testing. Sample 8001-D01 was introduced into the anaerobic chamber. The Eswab sample was easily vortexed. After suction, 50 μL of the sample was taken out. The sample aliquots were then centrifuged and pelletized. The pellet was prepared. DNA was extracted from the pellet, and the sample was evaluated by qPCR. The identity of the species was determined.

[0112] An additional 100 μL of sample was taken from the original Eswab sample. This aliquot Dilute it 10 times in a series, then 10 -3 , 10 -4 , 10 -5 Take 100 μL from the dilution. Spread the mixture onto MRS agar, then incubate at 35°C for 24-72 hours. Ta.

[0113] Lactobacillus species identified based on estimations, colony morphology, and Gram staining, then placed on an agar plate. Identified above. It has a colony morphology consistent with the estimated Lactobacillus colony morphology. Each individual colony is picked up using a sterile toothpick, and then placed on fresh MRS agar. Then, place on LAMVAB agar (a medium selective for Lactobacillus species), and then in a 96-degree deep wafer. The MRS broth in the wells of the plate was inoculated. Lactobacillus was grown in MRS broth. A 96-well plate containing Chillus was incubated for 24 hours. The rates were incubated at 35°C for 24-72 hours.

[0114] Strain 8001-D01-M006 was picked from one of the original MRS agar dilution plates. Raise grid spot #6 on MRS agar patch plate, on LAMVAB plate Spread over grid spot #6 and inoculated into MRS broth in well F7. 96 deep well The broth culture of strain 8001-D01-M006, grown in well F7 of the plate, was processed using 9 Well F7 of a 6-well PCR plate, and two 96-well microtiter plates Well F7 (contains 50% glycerol in the well, thereby 8001-D01) -When the M006 culture was added, the final glycerol concentration was 10%). 96 The culture, divided into smaller portions, is pelletized into well PCR plates, DNA is extracted, and the sample is processed using qP. The identity of Lactobacillus species was determined (where possible) by testing with CR. Strain 8001-D01-M006 was identified by qPCR as Lactobacillus gensenii ( It was determined to be Lactobacillus jensenii. Culture and 10 2 x 96 well microtiter plates containing % glycerol (freeze-preservation plates) The contents were sealed with adhesive foil seals and then frozen and stored at -70°C or below.

[0115] Stock 8001-D01-M006 was extracted from a frozen 96-well microtiter plate. The culture was harvested for propagation and characterization. One sample containing 10% glycerol was collected. A 96-well microtiter plate is placed in an anaerobic chamber and thawed at room temperature for 1 hour. The cultures in well F7 were gently mixed by pipetting, and then placed on MRS agar. Streaks were smeared for isolation. The plates were incubated at 35°C for 24–72 hours. .

[0116] Single-well isolated colonies were selected from streak plates and placed in pre-reduced MRS broth. It was used to inoculate the overnight culture (18-24 hours at 35°C). The next day, the overnight culture Dilute in fresh MRS broth and incubate for about 4 hours, then add 50% glycerin. Combine with cerol (final concentration of 10% glycerol), divide into small portions in freezer-safe vials ( Stored frozen at -70°C or below (1 ml per vial).

[0117] Strain 8001-D01-M006 was isolated, and pre-reduced MRS agar and broth were used. The culture medium was then propagated. The lot of the culture medium used was placed on the sample culture worksheet P249-01. Record information about 8001-D01. For each lot of MRS broth and MRS agar. Store the completed culture medium preparation worksheet in binder P249. Simply put, MR S agar and MRS broth growth medium containing the following components (gram weight / liter) H Ardy Diagnostic's Criterion® Lactobacillus MRS Prepared in-house using broth: dextrose (20.0g), pepsin digestion from animal tissue Ingredients (10.0g), beef extract (10.0g), yeast extract (5.0g), sodium acetate Um (5.0g), disodium phosphate (2.0g), ammonium citrate (2.0g) ), Tween 80 (1.0g), magnesium sulfate (0.1g), and manganese sulfate (0.05g). MRS agar plates use the same Criterion® Lactobacillus This product uses MRS broth as a base and contains 15 g / L of BD Difco agar. S broth and agar also contain 0.5 g / L of L-cysteine.

[0118] This strain, 8001-D01-M006, was detected by qPCR and Strainview sequencing. Identified by [method]. The qPCR method identifies amplicons generated from unknown isolates. By comparing the dissociation curve profile of the lactose with that of a known positive control strain, The genus Bacillus and seven different Lactobacillus species (Lactobacillus crispatus (Lc Lactobacillus rispatus, Lactobacillus gasseri, Lactobacillus Lactobacillus jensenii, Lactobacillus iners ), Lactobacillus acidophilus, Lactobacillus • Lactobacillus delbrueckii and Lactobacillus helvetii Detects L. helveticus. Strainview sequencing The project was carried out by Diversigen. The stock is Strainview Pipeline. When analyzed using the IV, it does not contain antibiotic resistance genes and does not contain pathogenicity genes. It was found that they do not possess it.

[0119] Strain 8001-D01-M006 is a suitable strain that can be used in vaginal microbiome compositions. Lactobacillus jensenii strain This could be one example.

[0120] Example 3 - Exemplary Lactobacillus gasseri (La) for use in a vaginal microbiome composition (Ctobacillus gasseri) strain Lactobacillus gasseri, designated as stock 8018-IN-M001. Donate (for example, as described herein) strains of *Illus gasseri*. The samples were isolated from donor specimens deposited (which were donated to the organization). Donors were collected using vaginal swabs and urine samples. Eligibility is obtained by donating a body, which is then subjected to Gram staining and Nugent scoring. Fairview Diagnostics Laboratory (for GRAM) It is sent to the next STI: Trichomonas vaginalis (TVPCR), Neisseria gonorrhoeae (GCPCR) ), Chlamydia trachomatis (CH I underwent a PCR test.

[0121] During the preparation of the 8018-IN-M001 working cell stock, the cultured cell density is set. Although it was not intended to be determined, a single well from a previously prepared image coating plate was used. By selecting isolated colonies, overnight culture was initiated. This overnight culture was then used the following morning. Dilute in fresh broth 1:5, allow to grow for at least 4 hours, then glycerinate the culture. Combined with the glycerol, a 10% glycerol stock was prepared. Generally, using this method... Adjust the glycerol stock to approximately 1 x 108 ~7×10 8 It has a cell density of CFU / ml This is providing a stock of resources.

[0122] OmniGene and ESwab vaginal swab specimens (8018-IN) were used for metagenomics. Sequencing (OmniGene), and culture and qPCR identity testing ( Provided for use as an ESwab. ESwab testing for culture and qPCR identity testing. Body 8018-IN was introduced into an anaerobic chamber. Eswab samples were easily vortexed. Then, 50 μL of the sample was taken out. The sample aliquots were pelletized by centrifugation. DNA was extracted from the pellet, and the sample was evaluated by qPCR to identify Lactobacillus species. This determined their identity.

[0123] An additional 100 μL of sample was taken from the original Eswab sample. This aliquot Dilute it 10 times in a series, then 10 -3 , 10 -4 , 10 -5 Take 100 μL from the dilution. Spread the mixture onto MRS agar, then incubate at 35°C for 24-72 hours. Ta.

[0124] Lactobacillus species identified based on estimations, colony morphology, and Gram staining, then placed on an agar plate. Identified above. It has a colony morphology consistent with the estimated Lactobacillus colony morphology. Each individual colony is picked up using a sterile toothpick, and then placed on fresh MRS agar. Then, place on LAMVAB agar (a medium selective for Lactobacillus species), and then in a 96-degree deep wafer. The MRS broth in the wells of the plate was inoculated. Lactobacillus was grown in MRS broth. A 96-well plate containing Chillus was incubated for 24 hours. The rates were incubated at 35°C for 24-72 hours.

[0125] Sampling of strain 8018-IN-M001 was picked up from one of the original MRS agar dilution plates. Okay, grid spot #1 on MRS agar patch plate, and grid spot #1 on LAMVAB plate. Spread over lid spot #1 and inoculated into MRS broth in well A7. 96 deep wells. The broth culture of strain 8018-IN-M001, which grows in well A7 of the plate, is 96 oz. Well A7 of a 96-well PCR plate, and two 96-well microtiter plates Well A7 (contains 50% glycerol in each well, thereby 8018-IN-M0 Transferred to 96 wells (at the time of adding the culture, the final glycerol concentration was 10%). The culture is pelletized in small portions on PCR plates, DNA is extracted, and the sample is subjected to qPCR. Therefore, tests were conducted to determine the identity of the Lactobacillus species (where possible). Strain 8 018-IN-M001 was identified by qPCR as Lactobacillus gasseri (Lactoba It was determined to be *Cillus gasseri*. Culture and 10% glycerol A 2x96 well microtiter plate (freezing plate) containing adhesive foil The contents were sealed with a seal and then frozen and stored at -70°C or below.

[0126] Stock 8018-IN-M001 was extracted from frozen 96-well microtiter plates. The culture was collected for propagation and characterization. One 9 oz container containing 10% glycerol was used. A 6-well microtiter plate was placed in an anaerobic chamber and thawed at room temperature for 1 hour. Gently mix the cultures in well A7 by pipetting, and then on MRS agar. Streaks were made for isolation. The plates were incubated at 35°C for 24–72 hours.

[0127] Single-well isolated colonies were selected from streak plates and placed in pre-reduced MRS broth. It was used to inoculate the overnight culture (18-24 hours at 35°C). The next day, the overnight culture Dilute in fresh MRS broth and incubate for about 4 hours, then add 50% glycerin. Combine with cerol (final concentration of 10% glycerol), divide into small portions in freezer-safe vials ( Stored frozen at -70°C or below (1 ml per vial).

[0128] Strain 8018-IN-M001 was isolated and prepared using pre-reduced MRS agar and broth. The culture medium was propagated. The lot of the culture medium used was placed on the P249-01 sample culture worksheet for sample 8. Record the details for 018-IN. Completion of each lot of MRS broth and MRS agar. Store the prepared culture medium worksheet in binder P249. In short, MRS cold The following components (grams / liter) of the Har dy Diagnostic's Criterion (trademark) Lactobacillus MRS blotch Prepared in-house using: dextrose (20.0g), pepsin digest of animal tissue (10.0g), beef extract (10.0g), yeast extract (5.0g), sodium acetate (5.0g), disodium phosphate (2.0g), ammonium citrate (2.0g), Tween 80 (1.0g), magnesium sulfate (0.1g), and manganese sulfate (0 (0.05g). MRS agar plates are the same Criterion (trademark) Lactobacillus genus. MRS broth is used as the base and contains 15 g / L of BD Difco agar. Ross and agar also contain 0.5 g / L of L-cysteine.

[0129] This strain 8018-IN-M001 was detected by qPCR and Strainview sequencing. Identified by [unclear]. The qPCR method identifies amplicons generated from unknown isolates. By comparing the dissociation curve profile of the lactobacillus with that of a known positive control strain, The genus Tylus and seven different Lactobacillus species (Lactobacillus crispatus (L. cr Lactobacillus ispatus, Lactobacillus gasseri, Lactobacillus Lactobacillus jensenii, Lactobacillus iners Lactobacillus acidophilus, Lactobacillus... Lactobacillus delbrueckii and Lactobacillus helvetii Detects L. helveticus. Strainview sequencing This was carried out by Diversigen. The strain is Strainview Pipeline When analyzed using this method, it is important to ensure that the sample does not contain antibiotic resistance genes and does not contain pathogenicity genes. It was found that this is not the case.

[0130] Strain 8018-IN-M001 is a suitable ra that can be used in vaginal microbiome compositions. This is an example of a Lactobacillus gasseri strain. obtain.

[0131] Example 4 - Exemplary Lactobacillus crispatus for use in vaginal microbiome compositions (Lactobacillus crispatus) strain Lactobacillus crispatus, designated as stock 8001-D01-M004 Donate (for example, as described herein) the strain of *Obacillus crispatus*. It was isolated from donor specimens deposited (as they were donated to be listed). The donor was vaginal Eligibility is obtained by donating wabs and urine samples, which are then Gram-stained and Nugen-stained. For t-score conversion (GRAM), Fairview Diagnostics Labo Sent to the uratory, next STI: Vaginal trichomoniasis (Trichomonas vag inalis) (TVPCR), Neisseria gonorrhoeae (GCPCR), Chlamydia trachomatis We performed a test for is(CHPCR).

[0132] During the preparation of the 8001-D01-M004 working cell stock, the cultured cell density was adjusted. Setting this was not targeted, but a single wave from a previously adjusted image coating plate By selecting isolated colonies, overnight culture was initiated. This overnight culture was then cultured the following morning. Dilute with fresh broth in a 1:5 ratio, let grow for at least 4 hours, then glycerinate the culture. Combined with the roll, a 10% glycerol stock was prepared. Generally, in this way... The adjustment of the glycerol stock is approximately 1 x 10 7 ~1 × 10 9 Cell density CFU / ml It brings with it the stocks it possesses.

[0133] OmniGene and ESwab vaginal swab specimens (8001-D01) were subjected to metagenomics. Sequencing (OmniGene), culture, and qPCR identity testing Provided for use as an ESwab. ESwab for culture and qPCR identity testing. Sample 8001-D01 was introduced into the anaerobic chamber. The Eswab sample was easily vortexed. After suction, 50 μL of the sample was taken out. The sample aliquots were then centrifuged and pelletized. The pellet was prepared. DNA was extracted from the pellet, and the sample was evaluated by qPCR. The identity of the species was determined.

[0134] An additional 100 μL of sample was taken from the original Eswab sample. This aliquot Dilute it 10 times in a series, then 10 -3 , 10 -4 , 10 -5 Take 100 μL from the dilution. Spread the mixture onto MRS agar, then incubate at 35°C for 24-72 hours. Ta.

[0135] Lactobacillus species identified based on estimations, colony morphology, and Gram staining, then placed on an agar plate. Identified above. It has a colony morphology consistent with the estimated Lactobacillus colony morphology. Each individual colony is picked up using a sterile toothpick, and then placed on fresh MRS agar. Then, place on LAMVAB agar (a medium selective for Lactobacillus species), and then in a 96-degree deep wafer. The MRS broth in the wells of the plate was inoculated. Lactobacillus was grown in MRS broth. A 96-well plate containing Chillus was incubated for 24 hours. The rates were incubated at 35°C for 24-72 hours.

[0136] Strain 8001-D01-M004 was picked from one of the original MRS agar dilution plates. Raise grid spot #4 on MRS agar patch plate, on LAMVAB plate Spread over grid spot #4 and inoculated into MRS broth in well D7. 96 deep well The broth culture of strain 8001-D01-M004, grown in well D7 of the plate, was processed in 9 Well D7 of a 6-well PCR plate, and two 96-well microtiter plates Well D7 (contains 50% glycerol in the well, thereby 8001-D01) -When the M004 culture was added, the final glycerol concentration was 10%). 96 The culture, divided into smaller portions, is pelletized into well PCR plates, DNA is extracted, and the sample is processed using qP. The identity of Lactobacillus species was determined (where possible) by testing with CR. Strain 8001-D01-M004 was identified by qPCR as Lactobacillus crispatus ( It was determined to be Lactobacillus crispatus. Culture and 1 2 x 96 well microtiter plates containing 0% glycerol (for cryopreservation) The contents were sealed with adhesive foil seals and then frozen and stored at -70°C or below.

[0137] Strain 8001-D01-M004 was extracted from a frozen 96-well microtiter plate. The culture was harvested for propagation and characterization. One sample containing 10% glycerol was collected. A 96-well microtiter plate is placed in an anaerobic chamber and thawed at room temperature for 1 hour. The cultures in well D7 were gently mixed by pipetting, and then placed on MRS agar. Streaks were smeared for isolation. The plates were incubated at 35°C for 24–72 hours. .

[0138] Single-well isolated colonies were selected from streak plates and placed in pre-reduced MRS broth. It was used to inoculate the overnight culture (18-24 hours at 35°C). The next day, the overnight culture Dilute in fresh MRS broth and incubate for about 4 hours, then add 50% glycerin. Combine with cerol (final concentration of 10% glycerol), divide into small portions in freezer-safe vials ( Stored frozen at -70°C or below (1 ml per vial).

[0139] Strain 8001-D01-M004 was isolated, and pre-reduced MRS agar and broth were used. The culture medium was then propagated. The lot of the culture medium used was placed on the sample culture worksheet P249-01. Record information about 8001-D01. For each lot of MRS broth and MRS agar. Store the completed culture medium preparation worksheet in binder P249. Simply put, MR S agar and MRS broth growth medium containing the following components (gram weight / liter) H Ardy Diagnostic's Criterion® Lactobacillus MRS Prepared in-house using broth: dextrose (20.0g), pepsin digestion from animal tissue Ingredients (10.0g), beef extract (10.0g), yeast extract (5.0g), sodium acetate Um (5.0g), disodium phosphate (2.0g), ammonium citrate (2.0g) ), Tween 80 (1.0g), magnesium sulfate (0.1g), and manganese sulfate (0.05g). MRS agar plates use the same Criterion® Lactobacillus This product uses MRS broth as a base and contains 15 g / L of BD Difco agar. S broth and agar also contain 0.5 g / L of L-cysteine.

[0140] This strain 8001-D01-M004 was found to be qPCR and Strainview sequencing. Identified by [method]. The qPCR method identifies amplicons generated from unknown isolates. By comparing the dissociation curve profile of the lactose with that of a known positive control strain, The genus Bacillus and seven different Lactobacillus species (Lactobacillus crispatus (Lc Lactobacillus rispatus, Lactobacillus gasseri, Lactobacillus Lactobacillus jensenii, Lactobacillus iners ), Lactobacillus acidophilus, Lactobacillus • Lactobacillus delbrueckii and Lactobacillus helvetii Detects L. helveticus. Strainview sequencing The project was carried out by Diversigen. The stock is Strainview Pipeline. When analyzed using the IV, it does not contain antibiotic resistance genes and does not contain pathogenicity genes. It was found that they do not possess it.

[0141] Strain 8001-D01-M004 is a suitable strain that can be used in vaginal microbiome compositions. Lactobacillus crispatus strain This could be one example.

[0142] Example 5 - Exemplary Lactobacillus rhamnosus for use in vaginal microbiome compositions (Lactobacillus rhamnosus) strain Lactobacillus rhamnosus, also known as strain BPL5 Exemplary strains of *Rhamnosus* are described in U.S. Patent No. 10,588,926. This is shown, and its entirety is incorporated herein by reference.

[0143] Strain BPL5 is a suitable Lactobacillus strain that can be used in vaginal microbiome compositions. This could be an example of a Lactobacillus rhamnosus strain.

[0144] Example 6 - pH-lowering ability of strain when grown anaerobically in MRS broth for 48 hours and milk Acid production Strain BPL5 (Lactobacillus rhamnosus; Example 6) , 8001-D01-M006 (Lactobacillus jensenii) );Example 2), 8001-D01-M004 (Lactobacillus crispatus (Lc rispatus); Example 4), and 8018-IN-M001 (Lactobacillus garnets). Water dropwort (L. gasseriri; Example 3) is used to lower the pH of MRS broth over time. Their in vitro capabilities were evaluated. To assess pH reduction, each strain was subjected to MRS blottery. The culture was inoculated into Ross and grown overnight (under anaerobic conditions). The next day, the overnight culture was reduced to approximately 1.0 OD 600nm (This is approximately 1 × 10 8 (Equivalent to CFU / mL) is adjusted, and then the new Further diluted (approximately 1 x 10) in bright MRS broth 6 The target starting concentration is CFU / mL. 8001-D01-M006, 8001-D01-M004, and 8018-IN-M0 Regarding 01): BPL5 strain was divided into two separate starting concentrations (approximately 1 × 10⁻⁶). 7 CFU / mL) The tests were conducted as follows: All cultures were incubated anaerobically for 48 hours, and the pH of the samples was determined as follows: Samples were taken at 0 and 48 hours of growth. All strains showed a significant change in the broth's pH by 48 hours. It can be reduced to Lactobacillus rhamnosus and The Lactobacillus crispatus strain exhibited the most significant pH reduction. The following is shown (Table 1). BPL5 (Lactobacillus rhamnosus (L. rhamnosus) sus)) and 8001-D01-M004 (Lactobacillus crispatus (L.cr Both strains of *ispatus* require the broth pH to remain below 4.0 for up to 48 hours. The values ​​were reduced (to 3.78 and 3.77, respectively). 8001-D01-M00 6 (Lactobacillus jensenii) and 8018-IN-M The 001 (Lactobacillus gasseri (L. gasseri)) strain also showed signs of aging within 48 hours. The loss pH was reduced (to 4.02 and 4.06, respectively), but Lactobacillus rhamno Lactobacillus rhamnosus and Lactobacillus crispatus It wasn't at the same level as tus.

[0145] [Table 1]

[0146] Additional samples were taken at 48 hours, filtered, and bacteria were removed. Then these The supernatant, free of cells, was analyzed to detect the levels of D-lactic acid and L-lactic acid. D- / L-Lactic Acid, produced by Megazyme, is derived from L-lactic acid. (D- / L-Lactate) Rapid Assay Kit (Catalog Number K-DLATE) Quantification was performed using lactate dehydrogenase and glutamate dehydrogenase. This kit uses lactate dehydrogenase and glutamate dehydrogenase. To quantify both D-lactic acid and L-lactic acid using an enzymatic reaction with genase. It is designed that the amount of NADH produced by the oxidation of lactic acid is equal to the starting concentration of lactic acid. It is stoichiometric. This kit was used according to the manufacturer's recommended procedure. Tested All strains produced detectable levels of lactic acid (Table 2). 8001-D01-M004 The Lactobacillus crispatus (L. crispus) strain has the highest level of D- It produces lactic acid and L-lactic acid (20.9 g / L) and the maximum amount of D-lactic acid (13.5 g), The BPL5 (Lactobacillus rhamnosus) strain has the largest amount It produced L-lactic acid (17.3 g / L) and a very small amount of D-lactic acid (0.8 g / L). Ctobacillus crispatus (L. crispatus) and Lactobacillus gasseri ( Both strains of L. gasseriri produce large amounts of both lactate isomers, and both strains are L-milk It produces more D-lactic acid than acid. Lactobacillus jensenii (L. jensenii) The 8001-D01-M006 strain of (nii) primarily produces D-lactic acid (12.4 g / L). This produced a very small amount of L-lactic acid (0.3 g / L).

[0147] [Table 2]

[0148] These results are from strain BPL5 (Lactobacillus rhamnosus (L. rhamnosus) );Example 6), 8001-D01-M006 (Lactobacillus gensenii (Lj ensenii); Example 2), 8001-D01-M004 (Lactobacillus Chris) Patas (L. crispatus); Example 4), and 8018-IN-M001 (L. Tobacillus gasseri (L. gasseri); Example 3) was used in the vaginal microbiome composition. This demonstrates that it can be a suitable material for that purpose.

[0149] Example 7 - Exemplary Vaginal Microbiota Composition An example of a vaginal microbiome composition is Lactobacillus crispatus (Lactobaci Lactobacillus crispatus, Lactobacillus gasseri Lactobacillus gasseri, Lactobacillus gensenii Lactobacillus jensenii, Lactobacillus rhamnosus It can be formed by combining (hamnosus). At least some examples So, Lactobacillus jensenii ) may be the strain described in Example 2. In at least some examples, lactobacill Lactobacillus gasseri is described in Example 3. It could be a strain. At least in some cases, Lactobacillus crispatus (Lactobacillus crispatus) Obacillus crispatus may be the strain described in Example 4. At least in some cases, Lactobacillus rhamnosus (Lactobacillus (s rhamnosus) may be the strain described in Example 5.

[0150] This disclosure should be understood to be illustrative in many respects. Changes in detail... In particular, without exceeding the scope of this disclosure, regarding the shape, size, and arrangement of the steps This can be done, to the extent appropriate, by the features of one exemplary embodiment. Any use of may include use in other embodiments. The scope of the present invention is naturally limited. However, the attached claims are defined in the language in which they are expressed.

Claims

1. A vaginal microbiome composition, Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus Lactobacillus jensenii, and Lactobacillus Bacteria including Lactobacillus rhamnosus A vaginal microbiome composition containing a mixture of the following.

2. The aforementioned bacterial mixture contains 25-75% Lactobacillus crispatus (Lactobacillus crispatus) The vaginal microbiome composition according to claim 1, comprising Cillus crispatus.

3. The aforementioned bacterial mixture contains 30-60% Lactobacillus crispatus (Lactobacillus crispatus) The vaginal cavity according to any one of claims 1 to 2, including Cillus crispatus. Microbiome composition.

4. The aforementioned bacterial mixture contains 40% Lactobacillus crispatus. Vaginal microorganisms according to any one of claims 1 to 3, including Lus crispatus. plexus composition.

5. The aforementioned bacterial mixture contains 1-10% Lactobacillus gasseri. Vaginal microbiome composition according to any one of claims 1 to 4, including (us gasseri) thing.

6. The aforementioned bacterial mixture contains 2-8% Lactobacillus gasseri A vaginal microbiome composition according to any one of claims 1 to 5, comprising (s gasser) 。

7. The aforementioned bacterial mixture contains 5% Lactobacillus gasseri. A vaginal microbiome composition according to any one of claims 1 to 6, comprising gasser.

8. The aforementioned bacterial mixture contains 1-10% Lactobacillus gensenii (Lactobacillus gensenii). The vaginal microorganism according to any one of claims 1 to 7, including Illus jensenii. Flora composition.

9. The aforementioned bacterial mixture contains 2-8% Lactobacillus gensenii. Vaginal microorganisms according to any one of claims 1 to 8, including (llus jensenii). plexus composition.

10. The aforementioned bacterial mixture contains 5% Lactobacillus gensenii. A vaginal microbiome group according to any one of claims 1 to 9, including (us jensenii). Finished product.

11. The aforementioned bacterial mixture contains 0.1-5% Lactobacillus acidophilus (Lactobacillus acidophilus). The following is a description of any one of claims 1 to 10, including acillus acidophilus. A composition of vaginal microbiota.

12. The aforementioned bacterial mixture contains 0.1-5% Lactobacillus rhamnosus (Lactobacillus rhamnosus). The vagina according to any one of claims 1 to 11, comprising cillus rhamnosus. Internal microbiota composition.

13. The aforementioned bacterial mixture contains 0.5-4% Lactobacillus rhamnosus (Lactobacillus rhamnosus). The vagina according to any one of claims 1 to 12, comprising cillus rhamnosus. Internal microbiota composition.

14. The aforementioned bacterial mixture contains 1% Lactobacillus rhamnosus (Lactobacillus rhamnosus). Vaginal microorganisms according to any one of claims 1 to 13, including Us rhamnosus. plexus composition.

15. The bacterial mixture is placed inside a capsule, as described in any one of claims 1 to 14. A composition of the vaginal microbiome.

16. The vaginal cavity according to any one of claims 1 to 15, wherein the bacterial mixture is placed in a suppository. Internal microbiota composition.

17. The bacterial mixture is disposed in a soluble shell, according to any one of claims 1 to 16. The described vaginal microbiome composition.

18. The aforementioned bacterial mixture is 1 × 10 ５ ~1 x 10 １５ Any of claims 1 to 16, including CFU A vaginal microbiome composition as described in any one of the items.

19. The aforementioned bacterial mixture is 1 × 10 ６ ~1 x 10 １２ Any of claims 1 to 16, including CFU A vaginal microbiome composition as described in any one of the items.

20. The aforementioned bacterial mixture is 1 × 10 ７ ~1 x 10 １０ Any of claims 1 to 16, including CFU A vaginal microbiome composition as described in any one of the items.

21. Any of claims 1 to 20, wherein the bacterial mixture comprises a freeze-dried bacterial mixture. The vaginal microbiome composition described in item 1.

22. A method for treating an infectious disease, wherein the method is The vaginal microbiome composition according to any one of claims 1 to 20 is administered to a patient with an infection. This includes giving, and the infection being bacterial vaginosis, candidiasis, human papillomavirus A method comprising one or more of the following: urinary tract infections, urinary tract infections, sexually transmitted infections, and gynecological cancers.

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