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Method for quickly detecting salmonella, shigella and Escherichia coli in faeces

A technology for Escherichia coli and Salmonella, applied in the field of biochemical detection, can solve problems such as difficult and rapid detection, and achieve the effects of reducing test workload, good application prospects and short detection time.

Inactive Publication Date: 2012-04-25
樊学军 +4
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The technical problem to be solved by the present invention is that it is difficult to rapidly detect and solve Salmonella, Shigella or Escherichia coli O157:H7 in feces samples in this field.

Method used

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  • Method for quickly detecting salmonella, shigella and Escherichia coli in faeces
  • Method for quickly detecting salmonella, shigella and Escherichia coli in faeces
  • Method for quickly detecting salmonella, shigella and Escherichia coli in faeces

Examples

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Effect test

Embodiment 1

[0038] Example 1 Preparation of Bare Ferrite Magnetic Beads and Optimization of Sample Pretreatment Conditions

[0039] In the present invention, a chemical precipitation method is used to prepare ferrite bare magnetic beads, and by optimizing and controlling reaction conditions, the distribution of magnetic particles is relatively uniform, and the size is about 10nm, reaching the nanometer level.

[0040] Weigh FeCl separately 2 4H 2 O and FeCl 3 ·6H 2 O, dissolved in deionized water, filtered and mixed; a small amount of 25% ammonia water is sprayed slowly and repeatedly, and the generated magnetic beads are small and fine, and the suspension time is long and stable. The prepared bare magnetic beads were stored under the environment of ammonia water, and were properly washed with sterilized deionized water before use.

[0041] Through the exploration of the adsorption conditions, it was determined that the best condition for the adsorption of the target bacteria by the b...

Embodiment 2

[0042] The establishment and optimization of embodiment two multiplex PCR system detection system

[0043] In the present invention, the target gene and the corresponding primers are screened by searching relevant reports at home and abroad, and its applicability to stool samples is explored through different combinations and optimizations. The genes selected for comparison include: uidA, rfbE conserved genes and virulence factor genes such as stx, eaeA, hly of Escherichia coli O157:H7, invA gene of Salmonella, and virulence factors such as ipaH, B, C, and D of Shigella factor gene.

[0044] Finally, the uidA and rfbE genes of Escherichia coli O157:H7, the invA gene of Salmonella, and the ipaH gene of Shigella were selected, and 5 pairs of primers were screened and designed using the conserved sequences of the target genes, and combined and matched to form different triplexes. The detection system was used to explore its sensitivity and specificity for detecting target bacter...

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Abstract

The invention aims to solve the technical problem that salmonella, shigella and Escherichia coli O157:H7 in a faeces sample are difficult to quickly detect and solve in the field of the prior art. In order to solve the technical problem, the technical scheme of the invention provides a multiple PCR (polymerase chain reaction) primer group and quick detection method for detecting the above bacteria among in-vitro bacteria. The multiple PCR primer group for detecting salmonella and shigella in in-vitro faeces contains a primer pair for salmonella invA genes and a primer pair for shigella ipaH genes. The invention also provides a method for carrying out multiple PCR amplification by using the primer group. Compared with the traditional method, the method provided by the invention has the characteristics of high speed, high accuracy, low cost and the like, can finish the detection of salmonella, shigella and Escherichia coli O157:H7 in faeces within 12 hours, and has good application prospects.

Description

technical field [0001] The invention belongs to the field of biochemical detection, in particular to a method for rapidly detecting Salmonella, Shigella and Escherichia coli in feces. Background technique [0002] Salmonella (Salmonella) is a common zoonotic intestinal pathogenic bacteria of great significance in public health, which can cause food poisoning, gastroenteritis, bacteremia, enteric fever and other diseases, so the World Health Organization (WHO) will Salmonella is listed as food-borne pathogens with serious and moderate hazards; Shigella is the pathogen that causes bacillary dysentery in humans, including Shigella dysenteriae, Shigella flexneri, Shigella baumannii and Shigella sonnei can cause food-borne outbreaks. The clinical manifestation is usually bloody diarrhea. It is mainly prevalent in developing countries, with more than 200 million cases worldwide every year; Escherichia coli O157:H7 (Escherichia coli O157:H7) is Escherichia coli that can cause hemo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12R1/42C12R1/19C12R1/01
Inventor 樊学军田绿波杨雨陈肖潇石莹
Owner 樊学军
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