CN104862370A - Bacteroides fragilis selective culture medium - Google Patents
Bacteroides fragilis selective culture medium Download PDFInfo
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- CN104862370A CN104862370A CN201510343214.3A CN201510343214A CN104862370A CN 104862370 A CN104862370 A CN 104862370A CN 201510343214 A CN201510343214 A CN 201510343214A CN 104862370 A CN104862370 A CN 104862370A
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Abstract
The invention discloses a bacteroides fragilis culture medium, and belongs to the field of inspection microbe cultivation. The bacteroides fragilis culture medium is characterized by comprising casein tryptone, glucose, yeast powder, sodium chloride, agar, cimigenyl xyloside, pig gallbladder powder, valine, and the balance of distilled water for per 1000 ml. Compared with the prior art, the bacteroides fragilis culture medium has the advantages that the inspection result is reliable, and the separation rate of bacteroides fragilis is high.
Description
Technical field
The present invention relates to Micro biological Tests field, be specifically related to a kind of bacteroides fragilis substratum.
Background technology
Bacteroides fragilis (Bacterooides Fragilis, BF) has pod membrane for Grain-negative tyrothricin, without brood cell, partly has pili, obligate anaerobic.BF is the integral part of Body normal flora, also be conditioned pathogen important clinically, the a lot of site infection of human body can be caused, infection symptoms is throughout clinical departments, recall rate is high, account for 25% of Clinical Anaerobic Bacteria strain isolated, genera bacillus strain isolated 50%, wherein diarrhea patient BF separation rate reaches 9.2% ~ 26.8%.
The clinical assays of BF is mainly PCR and cell cultures, and the susceptibility of PCR is high, within 5 hours, both can obtain result, and have quick and responsive characteristic, but there is following defect: 1. contain PCR inhibition in stool sample, easily cause false negative result; 2. complicated experiment condition is needed, costly.And cell cultures is the BF the most reliable method of inspection at present, but the obligate anaerobic of BF and severe matter of nourishing one's nature, cause its incubation time longer, when absolute anaerobic, use 2d to start have indivedual sample to start growth, the no positive bacterium colony of 7d occurs reaching negative test standard.Because qualification time is long, so above-mentioned inspection is difficult to guiding clinical treatment, miss an opportunity because of a delay.The substratum of the use that current BF cultivates mostly is bile Vitamin C2 (BBE) agar, its composition contains bile, bile in order to suppress to the anerobe of sensitivity, only have the bacteroides fragilis of resistance to bile and variable fusobacterium, fusobacterium mortiferum not suppressed, thus play the object of selectivity cultivation.But there is following defect in BBE substratum: the growth that 1. can not suppress variable fusobacterium, fusobacterium mortiferum; 2. for aerophil and facultative anaerobe insect killing effect not, therefore will add gentamicin 100mg/L, but gentamicin produces faint suppression operation equally for BF, causes its poor growth; If 3. do not add gentamicin, just need strict oxygen-free environment, because Clinical Laboratory In Grass Root Hospital lacks absolute anaerobic inspection machine mostly, therefore multiplex greatly " anaerobism cylinder culture method ", so-called anaerobism cylinder and dry cylinder, exhaust oxygen in cylinder by physicochemical method, cause anaerobic environment.The oxygen depletion of anaerobism cylinder method needs the time, the growth that have impact on obligate anaerobic bacillus BF be separated.
Therefore find a kind of new substratum, can under the environment of " cultivation of anaerobism cylinder " rapid detection, particularly important for basic unit's clinical hospitals.
Summary of the invention
Technical assignment of the present invention is for above the deficiencies in the prior art, provides a kind of Selective agar medium being suitable for the bacteroides fragilis growth of stool sample.
The technical scheme that the present invention solves its technical problem is: a kind of bacteroides fragilis Selective agar medium, is characterized in that, contains in often liter of substratum: pancreas casein peptone 10.0g; Glucose 2.0g; Yeast powder 5.0g; Sodium-chlor 5.0g; Agar 15.0g; Cimiside 3g; Pulvis Fellis Suis 20g; α-amino-isovaleric acid 0.1g; Aseptic calf serum 100ml; Distilled water adds to 1000ml.
The preparation method of above-mentioned substratum is: take pancreas casein peptone; Glucose; Yeast powder; Sodium-chlor; Agar; Cimiside; Pulvis Fellis Suis; α-amino-isovaleric acid, dissolve with distilled water, mix, sterilizing 15min at 121 DEG C, is cooled to 45 DEG C, adds the mixing of aseptic calf serum, packing after sterile purified water constant volume to 1000ml.
The present invention compared with prior art, has and detects the feature reliable, bacteroides fragilis separation rate is high.
1, described pancreas casein peptone contains various saccharides, amino acid, VITAMIN and trace element provides energy for bacteroides fragilis grows, glucose provides carbon source, the nutritive ingredients such as yeast powder rich in proteins, amino acid, polypeptide, Nucleotide, VITAMIN, somatomedin, trace element, promote the Growth and Reproduction of bacteroides fragilis; Sodium-chlor maintains balanced osmotic pressure; Agar is the peptizer of substratum;
2, α-amino-isovaleric acid is anerobe nutrition agent; L-homoarginine is activator, promotes the absorption of α-amino-isovaleric acid; γ-aminobutyric acid is nutrient intensifier, promotes bacteroides fragilis growth.
3, the use of Pulvis Fellis Suis and Cimiside (Cimicifugoside): the bile component in Pulvis Fellis Suis in order to suppress to the anerobe of sensitivity, and Cimiside is in order to be separated bacteroides fragilis and variable fusobacterium, fusobacterium mortiferum, its centrifugation is Selective depression anaerobism Fusobacterium nucleosides permeable membrane to transport, action pathway is outer membrane protein OMP on Fusobacterium cell walls and fadA adhesion protein, and because bacteroides fragilis is without above-mentioned specific proteins, therefore Cimiside for bacteroides fragilis without killing action.Infrastest confirms: bacteroides fragilis and variable fusobacterium, fusobacterium mortiferum bacterial strain are inoculated into respectively the Bile Agar substratum containing 3g/L Cimiside and common bile nutrient agar, in 37 DEG C of Anaerobic culturel 7d, bacteroides fragilis positive rate 100%, and variable fusobacterium, fusobacterium mortiferum bacterial strain positive rate are 0%.Confirm thus, Cimiside has good sterilization functions for variable fusobacterium, fusobacterium mortiferum, and has no significant effect for bacteroides fragilis.
4, thermal distortion calf serum has optionally antibacterial effect, and its thermal distortion method is: heat 10min at getting aseptic calf serum 80 DEG C, adds sterilizing trypsin digestion 90min at 45 DEG C, and the mass ratio of enzyme-to-substrate is 1:150.Common aseptic calf serum only plays the effect of nutraceutical matrix, there is no optionally antibacterial effect, and thermal distortion calf serum antibacterial properties is for aerophil, change aerophil cell wall structure on the one hand, cause permeability to increase and dead, the redox-potential of substratum can be reduced on the other hand, promotion oxygen-free environment and accelerate aerophil death, and this kind of anti-microbial effect is only directed to aerophil, then invalid for anerobe.By experiment the fusobacterium bacterial strain of same aerobic coli strain and anaerobism is inoculated into anaerobic culture medium respectively and with the addition of on the optimization anaerobic culture medium of the thermal distortion calf serum of 80ml/L in filling a prescription, cultivate at 37 DEG C, hermetically drying cylinder ring border, 24h, 48h identify, adding thermal distortion calf serum intestinal bacteria positive rate during 24h is 50%, and on the substratum not adding thermal distortion calf serum, intestinal bacteria positive rate is 80%; 48h detected result is add the substratum fusobacterium positive rate 70% of thermal distortion calf serum, fusobacterium positive rate 20% on the substratum not adding thermal distortion calf serum; Confirm that thermal distortion calf serum has good inhibition to aerophil in anaerobism cylinder thus, and promote that anerobe grows by accelerating oxygen-free environment.
5, Rhizome of Grass leaf Sweelflag (Acorus gramineus) belongs to Acoraceae, and be graminoid per nnial herb, medicinal effects is rhizome position.Modern medicine study shows, Rhizome of Grass leaf Sweelflag is containing compositions such as volatile oil and carbohydrate, organic acid, amino acid, inorganic elements such as α-trans-Isoasarones, there are the effects such as swelling and pain relieving, waking up the patient from unconsciousness by dissipating phlegm, ulcer mange, Rhizome of Grass leaf Sweelflag Aqueous extracts is good nutrition-fortifying agent, Rhizome of Grass leaf Sweelflag water logging agent (1:3), all has restraining effect in various degree to dermatophytess such as trichophyton, Trichophyton concentricum, star-shaped nocardias in vitro.Have report to point out, Rhizome of Grass leaf Sweelflag decoction has the effect of killing ascites cells.The research such as Masuda T finds that the α-trans-Isoasarone in Rhizome of Grass leaf Sweelflag all has restraining effect to streptococcus aureus, Candida albicans, intestinal bacteria etc.Experiment confirms, Rhizome of Grass leaf Sweelflag water cooking liquid under 200g/L concentration there is no killing action for anerobe, take anaerobic culture medium as contrast, to add 200g/L Rhizome of Grass leaf Sweelflag water cooking liquid in Anaerobic culturel based formulas for experimental group, inoculation bacteroides fragilis bacterial strain, the positive rate of 96h is respectively 70% and 60%, and the two compares no significant difference (P>0.05).
Embodiment
Below in conjunction with practical situation, the specific embodiment of the present invention is elaborated.
Embodiment 1, contains in the formula of preparation 1000ml substratum: pancreas casein peptone 10g; Glucose 2g; Yeast powder 5g; Sodium-chlor 5g; Agar 15g; Cimiside 3g; Pulvis Fellis Suis 20g; α-amino-isovaleric acid 0.1g; Aseptic calf serum 100ml; Distilled water adds to 1000ml.Preparation method: take pancreas casein peptone; Glucose; Yeast powder; Sodium-chlor; Agar; Cimiside; Pulvis Fellis Suis; α-amino-isovaleric acid, dissolve with distilled water, mix, sterilizing 15min at 121 DEG C, is cooled to 45 DEG C, adds the mixing of aseptic calf serum, packing after sterile purified water constant volume to 1000ml.
Aseptic calf serum in the present embodiment only plays the effect of nutraceutical matrix, there is no optionally antibacterial effect.
Embodiment 2, contains in the formula of preparation 1000ml substratum: pancreas casein peptone 8g; Glucose 5g; Yeast powder 8g; Sodium-chlor 5g; Agar 15g; Cimiside 2.5g; Pulvis Fellis Suis 20g; α-amino-isovaleric acid 0.3g; Aseptic thermal distortion calf serum 50ml; Distilled water adds to 1000ml.Preparation method: take pancreas casein peptone; Glucose; Yeast powder; Sodium-chlor; Agar; Cimiside; Pulvis Fellis Suis; α-amino-isovaleric acid; Dissolve with distilled water, mix, sterilizing 15min at 121 DEG C, is cooled to 45 DEG C, adds the mixing of aseptic thermal distortion calf serum, packing after sterile purified water constant volume to 1000ml.
Described aseptic thermal distortion calf serum preparation method heats 10min at getting aseptic calf serum 80 DEG C, adds sterilizing trypsin digestion 90min at 45 DEG C, and the mass ratio of enzyme-to-substrate is 1:150.Aseptic thermal distortion calf serum preparation method in other embodiments is in the same manner as in Example 1.
Embodiment 3, contains in the formula of preparation 1000ml substratum: pancreas casein peptone 10g; Glucose 3g; Yeast powder 10g; Sodium-chlor 5g; Agar 20g; Cimiside 3g; Pulvis Fellis Suis 15g; α-amino-isovaleric acid 0.5g; L-homoarginine 0.1g; Aseptic thermal distortion calf serum 80ml; Distilled water adds to 1000ml.Preparation method: take pancreas casein peptone; Glucose; Yeast powder; Sodium-chlor; Agar; Cimiside; Pulvis Fellis Suis; α-amino-isovaleric acid; L-homoarginine; Dissolve with distilled water, mix, sterilizing 15min at 121 DEG C, is cooled to 45 DEG C, adds the mixing of aseptic thermal distortion calf serum, packing after sterile purified water constant volume to 1000ml.
Embodiment 4, contains in the formula of preparation 1000ml substratum: pancreas casein peptone 5g; Glucose 5g; Yeast powder 10g; Sodium-chlor 5g; Agar 20g; Cimiside 2g; Pulvis Fellis Suis 20g; α-amino-isovaleric acid 0.3g; L-homoarginine 0.3g; Aseptic thermal distortion calf serum 80ml; The Rhizome of Grass leaf Sweelflag Aqueous extracts 250ml of concentration 200g/L; Distilled water adds to 1000ml.Preparation method: (1) is got 50g Rhizome of Grass leaf Sweelflag and added water boil 45min, filters, gets filtrate, and adjustment volume is 200g/L to concentration, obtains Rhizome of Grass leaf Sweelflag water cooking liquid 250ml; (2) pancreas casein peptone is taken; Glucose; Yeast powder; Sodium-chlor; Agar; Cimiside; Pulvis Fellis Suis; α-amino-isovaleric acid; L-homoarginine, dissolves mixing with distilled water, then adds in the Rhizoma Acori Gramineii extract liquid of step (1) gained and mixes 121 DEG C of sterilizing 15min, be cooled to 45 DEG C, adds the mixing of aseptic thermal distortion calf serum, packing after sterile purified water constant volume to 1000ml.
Embodiment 5, contains in the formula of preparation 1000ml substratum: pancreas casein peptone 8g; Glucose 3g; Yeast powder 10g; Sodium-chlor 5g; Agar 20g; Cimiside 3g; Pulvis Fellis Suis 15g; α-amino-isovaleric acid 0.3g; L-homoarginine 0.5g; γ-aminobutyric acid 0.1g; Aseptic thermal distortion calf serum 100ml; The Rhizome of Grass leaf Sweelflag Aqueous extracts 300ml of concentration 200g/L; Distilled water adds to 1000ml.Preparation method: preparation method: (1) is got 60g Rhizome of Grass leaf Sweelflag and added water boil 60min, filters, gets filtrate, and adjustment volume is 200g/L to concentration, obtains Rhizome of Grass leaf Sweelflag water cooking liquid 300ml; (2) pancreas casein peptone is taken; Glucose; Yeast powder; Sodium-chlor; Agar; Cimiside; Pulvis Fellis Suis; α-amino-isovaleric acid; L-homoarginine; γ-aminobutyric acid distilled water dissolves, mixing, and be dissolved in the Rhizoma Acori Gramineii extract liquid of step gained and mix, 121 DEG C of sterilizing 15min, are cooled to 45 DEG C, adds the mixing of aseptic thermal distortion calf serum, packing after sterile purified water constant volume to 1000ml.
Described Rhizome of Grass leaf Sweelflag Aqueous extracts concentration 200g/L refers to often liter containing crude drug 200g.
Gained bacteroides fragilis substratum of the present invention is used for stool sample, and have and detect the feature reliable, bacteroides fragilis separation rate is high, be clinical data sufficient proof, pertinent data is as follows.
1 object and method.
1.1 medium preparing: experimental group establishes 5 groups altogether, are respectively embodiment 1 scheme group, embodiment 2 scheme group, embodiment 3 scheme group, embodiment 4 scheme group, embodiment 5 scheme group.Control group is bile Vitamin C2 (BBE) agar purchased from Qingdao Hai Bo Bioisystech Co., Ltd, and wherein the concentration of bile is 20g/L, Vitamin C2 concentration is 1g/L.
1.2 collection of specimens cultural methods: 119 routine Diarrheas (3 ~ 12 years old age), are inoculated in each experimental group and control group substratum after gathering ight soil mucus.At 37 DEG C, anaerobism cylinder (common dry cylinder) is cultivated, and carries out identification of bacteria respectively at the tiny bacterium colony of 2d picking, as occurred carrying out identification of bacteria when being then cultured to 7d like bacterium colony undoubtedly.
1.3 detect positive criteria: according to " clinical microbiology diagnosis and diagram ", mode of appearance is bacterium colony canescence, circular dimpling, and smooth surface, neat in edge; Gram-negative; Indole reaction (-); Catalase reaction (+).
2 results: 19 routine Diarrhea stool samples cultivation 2d and the 7d positives detect situation and see the following form,
| Grouping | Sum | The positive number of cases of 2d | The positive number of cases of 7d |
| Embodiment 1 | 119 | 29 | 34 |
| Embodiment 2 | 119 | 31 | 34 |
| Embodiment 3 | 119 | 31 | 36 |
| Embodiment 4 | 119 | 33 | 35 |
| Embodiment 5 | 119 | 34 | 34 |
| Control group | 119 | 13 | 35 |
With BBE substratum 7d cultivation results for positive rate reference standard.The above results can be found out, after cultivating 2d, the positive rate of control group is starkly lower than standard value (P<0.001), show because anaerobism cylinder needs oxygen depletion process, can not provide a good anaerobic growing environment in 48h, miscellaneous bacteria breeding is rapid, bacteroides fragilis poor growth, therefore during 2d can not positive rate low, need 7d just can reach and detect object.And after each embodiment group cultivation 2d, positive rate compares with control group, all have notable difference (P<0.05), with standard value no significant difference (P>0.05), both each embodiment group 2d cultivation results had diagnostic value.
Substratum of the present invention is used for cultivating bacteroides fragilis under dry cylinder ring border, and detect reliable, separation rate is high, thus the time is made a definite diagnosis in shortening.
Claims (1)
1. a bacteroides fragilis Selective agar medium, is characterized in that, contains in often liter of substratum: pancreas casein peptone 10g; Glucose 2g; Yeast powder 5g; Sodium-chlor 5g; Agar 15g; Cimiside 3g; Pulvis Fellis Suis 20g; α-amino-isovaleric acid 0.1g; Aseptic calf serum 100ml; Distilled water adds to 1000ml.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104263807A (en) * | 2014-09-16 | 2015-01-07 | 青岛永通电梯工程有限公司 | Bile liquid culture medium and applications thereof |
| CN104651469A (en) * | 2013-11-25 | 2015-05-27 | 刘汉斌 | Bile liquid medium and use thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104651469A (en) * | 2013-11-25 | 2015-05-27 | 刘汉斌 | Bile liquid medium and use thereof |
| CN104263807A (en) * | 2014-09-16 | 2015-01-07 | 青岛永通电梯工程有限公司 | Bile liquid culture medium and applications thereof |
Non-Patent Citations (6)
| Title |
|---|
| 冯仁丰主编: "《实用医学检验学》", 31 December 1996 * |
| 刘永佳主编: "《临床医师手册 检验分册》", 31 December 1991 * |
| 杨彦 等: "龙胆泻肝丸(汤)治疗细菌性阴道病的临床与实验研究", 《医学理论与实践》 * |
| 王记祥 等: "南川升麻的化学成分和抗菌活性研究", 《安徽农业科学》 * |
| 胡桂清 等: "五种地产中药抗菌作用的实验研究", 《中医药学报》 * |
| 陈天寿主编: "《微生物培养基的制造与应用》", 30 June 1995 * |
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Application publication date: 20150826 |