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Microorganism ability verification salmonella sample preparation method

A Salmonella, proficiency testing technology, applied in biochemical equipment and methods, microbial determination/inspection, preservation of microorganisms, etc. Achieve the effect of high bacterial survival rate, reasonable standard of different difficulty levels, and good stability

Active Publication Date: 2015-12-23
舟山出入境检验检疫局综合技术服务中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at present, there is no high-reliability and high-throughput preparation technology for microbial proficiency testing in the field of microbiological testing, and there are many uncertain factors, which greatly reduces the credibility of inter-laboratory proficiency testing.
[0004] Cao Wenbo et al. disclosed a method for preparing samples for Salmonella proficiency testing in "Journal of Food Safety and Quality Inspection" in Volume 6, Issue 1, 2015: vacuum freeze-drying was used to prepare samples, which improved the quality of samples to a certain extent. Stability and uniformity, but still not ideal, and lack of difficulty level comparison standard
[0005] The existing Salmonella samples for microbial proficiency testing have poor strain stability and low survival rate, and the verification difficulty of different types of samples is different, and there is a lack of comparison standards, so the significance of microbial proficiency testing is also great. discount

Method used

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  • Microorganism ability verification salmonella sample preparation method

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] The preparation method of Salmonella sample for microbiological proficiency testing, with Salmonella as the target bacteria and Serratia marcescens, Citrobacter freundii, and Escherichia coli as interfering bacteria, includes the following steps:

[0034] (A), recovery and enrichment of bacterial strains: pick a ring of each bacterial strain with an inoculation loop, add the standard bacteria of the target bacteria and the standard bacteria of each interfering bacteria into 10mL of nutrient broth or nutrient agar slant medium respectively, The standard bacteria were revived, grown and enriched at 36±1° C., and the strains were identified during the culture period.

[0035] (B), isolation of bacterial strains: when the Salmonella cultured above is cultivated to the stationary phase, its culture medium is centrifuged to obtain the Salmonella bacteria sludge; when each interfering bacteria is cultivated to the logarithmic growth phase, its culture medium is carried out Cen...

Embodiment 2

[0046] The preparation method of Salmonella sample for microbiological proficiency testing, with Salmonella as the target bacteria and Serratia marcescens, Citrobacter freundii, and Escherichia coli as interfering bacteria, includes the following steps:

[0047] (A), recovery and enrichment of bacterial strains: pick a ring of each bacterial strain with an inoculation loop, add the standard bacteria of the target bacteria and the standard bacteria of each interfering bacteria into 10 mL of nutrient broth or nutrient agar slant medium respectively, The standard bacteria were revived, grown and enriched at 36±1° C., and the strains were identified during the culture period. Trehalose is added to the medium, and the content of the trehalose in the medium is 3 g / mL.

[0048] (B), isolation of bacterial strains: when the Salmonella cultured above is cultivated to a stable phase, the culture medium is placed in an environment of 48°C for 1.5h, and then the culture medium is centrifu...

Embodiment 3

[0059] The preparation method of Salmonella sample for microbiological proficiency testing, with Salmonella as the target bacteria and Serratia marcescens, Citrobacter freundii, and Escherichia coli as interfering bacteria, includes the following steps:

[0060] (A), recovery and enrichment of bacterial strains: pick a ring of each bacterial strain with an inoculation loop, add the standard bacteria of the target bacteria and the standard bacteria of each interfering bacteria into 10 mL of nutrient broth or nutrient agar slant medium respectively, The standard bacteria were revived, grown and enriched at 36±1° C., and the strains were identified during the culture period. Trehalose is added to the medium, and the content of the trehalose in the medium is 4 g / mL.

[0061] (B), isolation of bacterial strains: when the above-mentioned cultured Salmonella was cultivated to a stable stage, the culture medium was placed in an environment of 45° C. for 2 hours, and then the culture m...

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Abstract

The present invention relates to the technical field of microorganism ability verification, and discloses a microorganism ability verification salmonella sample preparation method. According to the method, salmonella is adopted as target bacteria, and Serratia marcescens, Citrobacter freundii and Escherichia coli are adopted as interference bacteria; the method comprises: (A) strain resuscitation and amplification; (B) strain isolation; (C) bacterial suspension preparation; and (D) freeze-drying and packaging; and the prepared samples comprise a simple-level negative sample, a simple-level positive sample, an intermediate-level negative sample, an intermediate-level positive sample, a difficult-level negative sample and a difficult-level positive sample, wherein the sample having the same level are subjected to paired use. According to the present invention, the sample prepared through the method has the good stability, the strain survival rate is high, and the reasonable level standards with different difficulties are provided.

Description

technical field [0001] The invention relates to the technical field of microbial proficiency verification, in particular to a method for preparing a Salmonella sample for microbial proficiency verification. Background technique [0002] Proficiency testing is one of the effective measures for laboratory external quality control, and it is also a necessary condition for metrological certification and laboratory accreditation. The implementation of microbiological proficiency testing can effectively evaluate the technical ability of participating laboratories to carry out relevant microbiological tests. [0003] Salmonella, as a common pathogenic bacteria in food poisoning and foodborne diseases in my country in recent years, is the target bacteria often used in proficiency testing projects. However, at present, there is no high-reliability and high-throughput microbial proficiency testing preparation technology in the field of microbiology testing, and there are many uncerta...

Claims

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Application Information

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IPC IPC(8): C12N1/04C12N1/20C12Q1/02
Inventor 胡兴娟沈飚邵宏宏周秀锦徐君辉杨赛军
Owner 舟山出入境检验检疫局综合技术服务中心
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