CN104694434A - Culture medium capable of simultaneously proliferating multiple pathogenic bacteria in meat products and preparation method of culture medium - Google Patents
Culture medium capable of simultaneously proliferating multiple pathogenic bacteria in meat products and preparation method of culture medium Download PDFInfo
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- CN104694434A CN104694434A CN201510110525.5A CN201510110525A CN104694434A CN 104694434 A CN104694434 A CN 104694434A CN 201510110525 A CN201510110525 A CN 201510110525A CN 104694434 A CN104694434 A CN 104694434A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention relates to a culture medium capable of simultaneously proliferating multiple pathogenic bacteria in meat products and a preparation method of the culture medium. The culture medium comprises the following components: 20.0g/L of peptone, 10.0g/L of sodium chloride, 18.0g/L of disodium hydrogen phosphate (Na2HPO4.12H2O) and 3.0g/L of monopotassium phosphate. The preparation method comprises the following steps: mixing, heating and stirring the previous components until the components are completely dissolved, cooling to room temperature, regulating the pH value to be 7.2+/-0.1, and performing autoclaved sterilization at the temperature of 121 DEG C for 15 minutes. The culture medium can simultaneously perform composite enrichment on salmonella, staphylococcus aureus, listeria monocytogenes and diarrheogenic Escherichia coli, the complicated process of respective enrichment is avoided, the working efficiency is improved, and the culture medium has significance on quarantine inspection of pathogenic bacteria in meat products.
Description
Technical field
The present invention relates to a kind of bacteria culture medium, particularly relate to and a kind ofly can breed substratum of various pathogens in meat product and preparation method thereof simultaneously.
Background technology
At present, pathogenic microbes detect is all generally carry out according to related standards method, and first standard method will increase bacterium respectively, and namely a kind of bacterium carries out increasing bacterium according to a set of enrichment medium.Pathogenic microbes detect generally will detect various pathogens simultaneously, if each pathogenic bacterium adopts a set of enrichment medium, testing can be caused loaded down with trivial details.
Table 1 GB increases bacterium method and increases comparing of bacterium method with compound
| Detection method | Detect the time limit | Sensitivity | Operability | Genome extracts |
| GB increases bacterium method | 6～24h | 10～3000 | Loaded down with trivial details (increasing bacterium 6 times) | Extract 4 |
| Compound enrichment | 6～24h | 10～2000 | Simply (increase bacterium 1 time) | Extract 1 |
Summary of the invention
Based on above weak point, the invention provides and a kind ofly can breed substratum of various pathogens in meat product and preparation method thereof simultaneously, can carry out to various pathogens such as Salmonellas, streptococcus aureus, Listeria Monocytogenes, Diarrheogenil Escherichia colis the cultivation increasing bacterium simultaneously, improve the efficiency increasing bacterium.
Object of the present invention is realized by following technology:
1, can breed a substratum for various pathogens in meat product, medium component is simultaneously: peptone 20.0g/L, sodium-chlor 10.0g/L, Sodium phosphate dibasic (Na
2hPO
412H
2o) 18.0g/L and potassium primary phosphate 3.0g/L.
The present invention also has following technical characteristic:
1, a kind of substratum simultaneously breeding various pathogens in meat product as above, compound method is as follows:
(1) join in distilled water by peptone, sodium-chlor, Sodium phosphate dibasic, potassium primary phosphate, heated and stirred is to dissolving completely;
(2), after being cooled to room temperature, pH value to 7.2 ± 0.1 is adjusted;
(3) 121 DEG C of autoclaving 15min; Cooling.
2, a kind of substratum simultaneously breeding various pathogens in meat product as above, the application in the quarantine of meat product pathogenic bacterium.
Beneficial effect of the present invention is embodied in: basal culture medium can carry out compound increase bacterium to Salmonellas, streptococcus aureus, Listeria Monocytogenes, Diarrheogenil Escherichia coli simultaneously, avoid the complicated procedures increasing bacterium separately, improve working efficiency; The culture increased after bacterium can carry out separation and Culture and bioassay experiments, also can be directly used in PCR and detect.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
embodiment 1
The selection of compound enrichment medium
According to the growth characteristics of pathogenic bacterium in meat product, species variation, searches compound enrichment medium, and needs the features such as reparation in conjunction with pathogenic bacterium subject to damage in food, filters out two kinds of substratum.Two kinds of substratum Zengjing Granule Salmonellas, streptococcus aureus, Listeria Monocytogenes, Diarrheogenil Escherichia coli respectively.According to increasing bacterium situation analysis total number of bacterial colony and incubation time, determine that the substratum increasing bacterium efficiency high is optimum medium.
embodiment 2
The establishment of compound Zengjing Granule based component and compound method thereof
1, the establishment of compound Zengjing Granule based component
Medium component is: peptone 20.0g, sodium-chlor 10.0g, Sodium phosphate dibasic (Na
2hPO
412H
2o) 18.0g, potassium primary phosphate 3.0g, distilled water 1000mL.
2, the preparation method of compound enrichment medium
Concrete preparation method and cultural method as follows:
(1) join in distilled water by peptone, sodium-chlor, Sodium phosphate dibasic, potassium primary phosphate, heated and stirred is to dissolving completely;
(2), after being cooled to room temperature, pH value to 7.2 ± 0.1 is adjusted;
(3) 121 DEG C of autoclaving 15min;
(4), after substratum cooling, 18 ~ 24h can be cultivated at 36 DEG C.
embodiment 3
The application of compound enrichment medium
1, with compound enrichment medium dilute Salmonellas respectively, streptococcus aureus, monocyte increase bacterium listeria spp, the nutrient solution of pathogenic colon bacillus, enterotoxigenic Escherichia coli, hemorrhagic colon bacillus O157:H7, enteron aisle aggressive colon bacillus to finite concentration, count.Then 36 DEG C of cultivation 24h count again, and compound enriching effect is in table 2.Visible, increase bacterium through compound and can meet detection demand.
Table 2 pathogenic bacterium compound enriching effect (unit: CFU/mL)
| Bacterial strain | Total number of bacterial colony before increasing bacterium | Total number of bacterial colony after increasing bacterium |
| Salmonellas | 50 | 5800000 |
| EHEC O157：H7 | 60 | 610000 |
| Streptococcus aureus | 90 | 87000 |
| Pathogenic colon bacillus | 70 | 640000 |
| Enterotoxigenic Escherichia coli | 56 | 530000 |
| Monocyte increases bacterium listeria spp | 70 | 710000 |
| Enteron aisle aggressive colon bacillus | 76 | 730000 |
2, meat product 25g is got with aseptic technique, add in the aseptic triangular flask or slap type homogenizing bag that 225mL complex medium is housed after pulverizing, Salmonellas, streptococcus aureus, monocyte increasing bacterium listeria spp, hemorrhagic colon bacillus O157:H7 is vaccinated with in complex medium, rock 3 ~ 5min or bounce 1min, cultivating 6h by after triangular flask or homogenizing bag sealing at 36 DEG C.From substratum, take out 10 mL joins in the test tube that 10mL complex medium is housed, and cultivates 24h after mixing at 36 DEG C.From complex medium, extract the nucleic acid of pathogenic bacterium, carry out the PCR detection of Salmonellas, streptococcus aureus, monocyte increasing bacterium listeria spp, hemorrhagic colon bacillus O157:H7, the positive bacteria of result inoculation all detects.
Claims (3)
1. can breed a substratum for various pathogens in meat product, it is characterized in that, medium component is simultaneously: peptone 20.0g/L, sodium-chlor 10.0g/L, Sodium phosphate dibasic (Na
2hPO
412H
2o) 18.0g/L and potassium primary phosphate 3.0g/L.
2. a kind of substratum simultaneously breeding various pathogens in meat product according to claim 1, it is characterized in that, compound method is as follows:
(1) by peptone, sodium-chlor, Sodium phosphate dibasic (Na
2hPO
412H
2o), potassium primary phosphate joins in distilled water, and heated and stirred is to dissolving completely;
(2), after being cooled to room temperature, pH=7.2 ± 0.1 is adjusted;
(3) 121 DEG C of autoclaving 15min; Cooling.
3. a kind of substratum simultaneously breeding various pathogens in meat product according to claim 1, is characterized in that, the application in the quarantine of meat product pathogenic bacterium.
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| CN201510110525.5A CN104694434A (en) | 2015-03-13 | 2015-03-13 | Culture medium capable of simultaneously proliferating multiple pathogenic bacteria in meat products and preparation method of culture medium |
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| CN201510110525.5A CN104694434A (en) | 2015-03-13 | 2015-03-13 | Culture medium capable of simultaneously proliferating multiple pathogenic bacteria in meat products and preparation method of culture medium |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101045909A (en) * | 2006-03-27 | 2007-10-03 | 陈福生 | Culture medium with simultaneoulsy enriched salmonella, colibacillus and Staphylococcus aureus |
| CN102660474A (en) * | 2012-05-05 | 2012-09-12 | 西南民族大学 | Culture medium capable of simultaneously enriching five kinds of food-borne pathogenic bacteria and preparation method for culture medium |
-
2015
- 2015-03-13 CN CN201510110525.5A patent/CN104694434A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101045909A (en) * | 2006-03-27 | 2007-10-03 | 陈福生 | Culture medium with simultaneoulsy enriched salmonella, colibacillus and Staphylococcus aureus |
| CN102660474A (en) * | 2012-05-05 | 2012-09-12 | 西南民族大学 | Culture medium capable of simultaneously enriching five kinds of food-borne pathogenic bacteria and preparation method for culture medium |
Non-Patent Citations (6)
| Title |
|---|
| HYOCHIN KIM, ET AL.: "SEL, a Selective Enrichment Broth for Simultaneous Growth of Salmonella enterica, Escherichia coli O157:H7, and Listeria monocytogenes", 《APPLIED AND ENVIRONMENTAL MICROBIOLOGY》 * |
| 凌代文 主编: "《乳酸细菌分类鉴定及实验方法》", 31 March 1999 * |
| 国家认证认可监督管理委员会 编: "《出入境检验检疫行业标准汇编 食品、化妆品检验卷》", 31 August 2012 * |
| 徐义刚 等: "多重PCR-DHPLC 检测4 种主要致腹泻性大肠埃希氏菌方法的建立与应用", 《食品科学》 * |
| 杨秀伟 等主编: "《中药成分代谢分析》", 30 September 2003, 中国医药科技出版社 * |
| 钱毅 赵国君 译: "《食品分析法》", 30 November 1990, 上海科学普及出版社 * |
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Application publication date: 20150610 |