CN107043705A - Prebiotic bacterial screening method for setting up clinical nutrition microorganism resource storehouse - Google Patents
Prebiotic bacterial screening method for setting up clinical nutrition microorganism resource storehouse Download PDFInfo
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Abstract
The present invention discloses a kind of prebiotic bacterial screening method for being used to set up clinical nutrition microorganism resource storehouse, is that the directional collecting system based on function of setting up improves just screen capacity first；And then suitable stool sample transport preservation system is established, extend 2 ~ 4 times of effective shipping time of stool sample, it is ensured that the probiotics strain quantity of separation survival；It is last to be filtered out respectively with the excellent species for promoting antibacterial and biological antagonist function, promotion metabolic function and promotion immunologic function function according to specific optimization test method, simple to operate, screening efficiency is high, available for setting up clinical nutrition microorganism resource storehouse, selected to carry out probiotics compounding for individuation difference, improve therapeutic effect.
Description
Technical field
The present invention relates to a kind of prebiotic bacterial screening method, especially a kind of simple to operate, screening efficiency it is high be used for set up
The prebiotic bacterial screening method in clinical nutrition microorganism resource storehouse.
Background technology
Probiotics is to be colonized in human body intestinal canal, in reproductive system, can produce definite health efficacy to improve the micro- life of host
State balance, the active beneficial microorganism general name for playing beneficial effect.Beneficial bacterium or fungi are main in human body, animal body
Have：Clostridium butyricum, lactobacillus, Bifidobacterium, actinomyces, saccharomycete, bacillus subtilis, Lactobacillus plantarum etc..Probiotics
Medicable disease is more, such as participates in the synthesis of vitamin, participates in cholesterol metabolic, and regulation cholesterol is diseases related；Preventing and treating
Allergy（Food hypersenstivity, asthma, rhinitis）, prevent eczema；Treat antibiotic associated diarrhea, alleviate IBS, IBD, bacterial vaginosis
Inflammation, urogenital infections, necrotizing enterocolitis；Control hypertension, regulation central nervous system function and obstacle；It is anti-deep and remote
Door pylori infection；Improve immune resistance infection；Also it can adjust lactose intolerance etc..Because single strain function has limitation
Property, therefore the composite reactive probiotics agents treatment disease effects of existing document report each quasi-microorganism compounding by more than are more preferable.
But, screening technique complexity, the efficiency of existing probiotics are low, therefore the bacterial strain that can be really used for clinical and nutrition at present is limited,
It is difficult to set up clinical nutrition microorganism resource storehouse, the no rationale of clinic compounding, it is impossible to be adjusted, lead for individuation difference
Cause therapeutic effect not ideal enough.
The content of the invention
The present invention is that there is provided a kind of simple to operate, screening effect in order to solve the above-mentioned technical problem present in prior art
The high prebiotic bacterial screening method for being used to set up clinical nutrition microorganism resource storehouse of rate.
The present invention technical solution be：A kind of probiotics for being used to set up clinical nutrition microorganism resource storehouse
Screening technique, it is characterised in that carry out in accordance with the following steps successively：
A. crowd's fresh excreta that collection metabolic function is good respectively, immunity is strong and anti-infection ability is strong is sample；
B. various probiotics strains are filtered out with selective medium；
C. further filtered out with antibacterial, biological antagonist function the excellent probiotics strain of promotion, with promotion by experiment
The excellent probiotics strain of metabolic function and the excellent probiotics strain for promoting immunologic function：
C.1. screened according to basic biological nature
Test the adhesive capacity of the tolerance of sour environment and bile, bacterium, by tolerance is good, adhesive capacity is strong
Bacterial strain be used as preliminary screening bacterial strain carry out next step screening；
C.2. colonization resistance function in-vitro screening
Bacteriostatic experiment is carried out to the bacterial strain of preliminary screening, screening can effectively suppress the probiotics strain of germ, and the germ is big
The uncommon bacterium of intestines angstrom, staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans；
C.3. metabolic function in-vitro screening is promoted
By trypsase, pepsin, extracellular protease, peptase, alcohol dehydrogenase, the acetaldehyde of the bacterial strain for detecting preliminary screening
Dehydrogenase isoreactivity, probiotics strain of the screening with different metabolic function；
C.4 immunologic function in-vitro screening is promoted
The experimental cell screened using PBMC as ion vitro immunization, detects shadow of the bacterial strain to PBMC cytokine secretions of preliminary screening
Ring, the strong probiotics strain of screening immunostimulation function.
Probiotic bacteria culture medium and sheep blood are added after a steps collect specimen into sample, 4 DEG C maintain up to 4 days.
The present invention is that the directional collecting system based on function of setting up improves just screen capacity first；And then establish suitable excrement mark
This transport preservation system, extends 2 ~ 4 times of effective shipping time of stool sample, it is ensured that the probiotics strain number of separation survival
Amount；It is last to be filtered out respectively with the antibacterial and biological antagonist function of promotion according to specific optimization test method, promote metabolism work(
The excellent species of immunologic function function and can be promoted, simple to operate, screening efficiency is high, available for setting up clinical nutrition microorganism resource
Storehouse, selects to carry out probiotics compounding for individuation difference, improves therapeutic effect.
Embodiment
A kind of prebiotic bacterial screening method for being used to set up clinical nutrition microorganism resource storehouse, is carried out in accordance with the following steps successively：
A. crowd's fresh excreta that collection metabolic function is good respectively, immunity is strong and anti-infection ability is strong is sample；Such as collection
Sample is needed to transport for a long time, then probiotic bacteria culture medium and sheep blood, 4 DEG C are added into sample
Maintain up to 4 days；
B. various probiotics strains are filtered out with selective medium；
C. further filtered out with antibacterial, biological antagonist function the excellent probiotics strain of promotion, with promotion by experiment
The excellent probiotics strain of metabolic function and the excellent probiotics strain for promoting immunologic function：
C.1. screened according to basic biological nature
Test the adhesive capacity of the tolerance of sour environment and bile, bacterium, by tolerance is good, adhesive capacity is strong
Bacterial strain be used as preliminary screening bacterial strain carry out next step screening；
C.2. colonization resistance function in-vitro screening
Bacteriostatic experiment is carried out to the bacterial strain of preliminary screening, screening can effectively suppress the probiotics strain of germ, and the germ is big
The uncommon bacterium of intestines angstrom, staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans；
C.3. metabolic function in-vitro screening system is promoted
By trypsase, pepsin, extracellular protease, peptase, alcohol dehydrogenase, the acetaldehyde of the bacterial strain for detecting preliminary screening
Dehydrogenase isoreactivity, probiotics strain of the screening with different metabolic function；
C.4 immunologic function in-vitro screening system is promoted
With the PBMC for being relatively easy to be separately cultured and being reacted with preferable immunostimulation（PMNC）As external
The experimental cell of immunoscreening, detects influence of the bacterial strain of preliminary screening to PBMC cytokine secretions, screens immunostimulation work(
The strong probiotics strain of energy.
Experiment：
Acid resistance ability is low and the high bacterial strain of acid resistance ability is divided into two groups, respectively to EHEC and staphylococcus aureus
Bacteriostatic experiment is carried out, acid resistance ability is low and the high bacterial strain bacteriostasis of acid resistance ability has marked difference（P <0.05）.It is acidproof
Sexuality is related to bacteriostasis（EHEC R=0.465, p<0.05；Staphylococcus aureus R=0.384, p<0.05）.
It therefore, it can go out the probiotics with certain bacteriostasis by c.1 preliminary screening.
Claims (2)
1. a kind of prebiotic bacterial screening method for being used to set up clinical nutrition microorganism resource storehouse, it is characterised in that successively
Carry out in accordance with the following steps：
A. crowd's fresh excreta that collection metabolic function is good respectively, immunity is strong and anti-infection ability is strong is sample；
B. various probiotics strains are filtered out with selective medium；
C. further filtered out with antibacterial, biological antagonist function the excellent probiotics strain of promotion, with promotion by experiment
The excellent probiotics strain of metabolic function and the excellent probiotics strain for promoting immunologic function：
C.1 screened according to basic biological nature
Test the adhesive capacity of the tolerance of sour environment and bile, bacterium, by tolerance is good, adhesive capacity is strong
Bacterial strain be used as preliminary screening bacterial strain carry out next step screening；
C.2 colonization resistance function in-vitro screening
Bacteriostatic experiment is carried out to the bacterial strain of preliminary screening, screening can effectively suppress the probiotics strain of germ, and the germ is big
The uncommon bacterium of intestines angstrom, staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans；
C.3 metabolic function in-vitro screening is promoted
By trypsase, pepsin, extracellular protease, peptase, alcohol dehydrogenase, the acetaldehyde of the bacterial strain for detecting preliminary screening
Dehydrogenase isoreactivity, probiotics strain of the screening with different metabolic function；
C.4 immunologic function in-vitro screening is promoted
The experimental cell screened using PBMC as ion vitro immunization, detects shadow of the bacterial strain to PBMC cytokine secretions of preliminary screening
Ring, the strong probiotics strain of screening immunostimulation function.
2. the prebiotic bacterial screening method according to claim 1 for being used to set up clinical nutrition microorganism resource storehouse, its feature exists
In：Probiotic bacteria culture medium and sheep blood are added after a steps collect specimen into sample, 4 DEG C maintain up to 4 days.
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000041707A2 (en) * | 1999-01-15 | 2000-07-20 | Enterprise Ireland (Trading As Bioresearch Ireland) | USE OF $i(LACTOBACILLUS SALIVARIUS) |
| CN101260377A (en) * | 2008-03-14 | 2008-09-10 | 内蒙古蒙牛乳业（集团）股份有限公司 | Animal bifidobacteria and use thereof |
| CN101649303A (en) * | 2009-05-08 | 2010-02-17 | 内蒙古蒙牛乳业（集团）股份有限公司 | Bifidobacterium, application thereof and food composition containing same |
| CN101688171A (en) * | 2007-03-28 | 2010-03-31 | 营养健康有限公司 | probiotic bifidobacterium strains |
| CN102373172A (en) * | 2011-11-03 | 2012-03-14 | 北京龙科方舟生物工程技术中心 | Enterococcus faecium and application thereof |
| CN102864091A (en) * | 2012-01-15 | 2013-01-09 | 河南科技大学 | One-bacterium multiple-enzyme bacterial strain as well as screening method and application thereof |
| CN105112333A (en) * | 2015-08-31 | 2015-12-02 | 江南大学 | Bifidobacterium longum with good intestinal tract colonizing ability and screening method and application of bifidobacterium longum |
-
2016
- 2016-12-30 CN CN201611263098.5A patent/CN107043705A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2000041707A2 (en) * | 1999-01-15 | 2000-07-20 | Enterprise Ireland (Trading As Bioresearch Ireland) | USE OF $i(LACTOBACILLUS SALIVARIUS) |
| CN101688171A (en) * | 2007-03-28 | 2010-03-31 | 营养健康有限公司 | probiotic bifidobacterium strains |
| CN101260377A (en) * | 2008-03-14 | 2008-09-10 | 内蒙古蒙牛乳业（集团）股份有限公司 | Animal bifidobacteria and use thereof |
| CN101649303A (en) * | 2009-05-08 | 2010-02-17 | 内蒙古蒙牛乳业（集团）股份有限公司 | Bifidobacterium, application thereof and food composition containing same |
| CN102373172A (en) * | 2011-11-03 | 2012-03-14 | 北京龙科方舟生物工程技术中心 | Enterococcus faecium and application thereof |
| CN102864091A (en) * | 2012-01-15 | 2013-01-09 | 河南科技大学 | One-bacterium multiple-enzyme bacterial strain as well as screening method and application thereof |
| CN105112333A (en) * | 2015-08-31 | 2015-12-02 | 江南大学 | Bifidobacterium longum with good intestinal tract colonizing ability and screening method and application of bifidobacterium longum |
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