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Quick high-throughput intestines source pathogenic bacterium detection method

A technology of intestinal pathogenic bacteria and detection method, which is applied in the field of in vitro diagnostic reagents, can solve problems such as complicated operation, and achieve the effect of improving detection ability, reducing time and complexity, and fast and accurate detection

Inactive Publication Date: 2014-07-02
SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] At present, gene chip detection for pathogenic bacteria detection has been used in some researches, but the probes of these gene chips are mostly placed in a manual mode, which is complicated to operate and increases the chance of many false positives

Method used

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  • Quick high-throughput intestines source pathogenic bacterium detection method
  • Quick high-throughput intestines source pathogenic bacterium detection method
  • Quick high-throughput intestines source pathogenic bacterium detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0079] The specificity and sensitivity of the detection of five kinds of enterogenous pathogenic bacteria of embodiment 1

[0080] 1. Experimental steps

[0081] 1. Recovery of Bacteria

[0082] Inoculate Shigella fusiformis, Salmonella, Vibrio parahaemolyticus, Staphylococcus aureus, and Listeria monocytogenes in nutrient broth medium, culture at 37°C for 12 hours, streak inoculate on nutrient agar plate, and culture at 37°C After 24 hours, a single colony was picked and cultured in LB medium at 37°C for 18 hours.

[0083] 2. Identification of standard strains

[0084] Standard strains of Shigella fumigatus, Salmonella, Vibrio parahaemolyticus, and Listeria monocytogenes were purchased from ATCC in the United States, and Staphylococcus aureus was a positive strain isolated from the laboratory.

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Abstract

The invention relates to a quick, sensitive and high-throughput intestines source pathogenic bacterium detection method. The detection method provided by the invention integrates two powerful molecular biological techniques: polymerase chain reaction PCR and a micro-array, and directly fixes a probe of PCR hybridization in a hybridization cabin of the micro-array on a same chip with a PCR reaction chamber. The detection method comprises the following steps of enriching bacteria; extracting a DNA solution; carrying out PCR amplification; hybridizing; cleaning; and judging the result. The method provided by the invention can quickly detect genes of vibrio parahaemolyticus, Shigella, staphylococcus aureus, listeria monocytogenes and salmonella in high throughput, and the detection efficiency of front-line inspection and quarantine personnel of import and export ports can be greatly improved, thereby not only reducing the workload, but also solving the undetected positive result problem probably caused by conventional detection method to the maximum extent. Therefore, food safety incidents are prevented to the maximum extent.

Description

technical field [0001] The invention relates to the technical field of in vitro diagnostic reagents, in particular to a rapid and high-throughput detection method for intestinal pathogenic bacteria. Background technique [0002] Infectious diarrhea caused by foodborne pathogens is the second most common cause of morbidity and mortality worldwide after cardiovascular disease. Judging from the incidence of legally notifiable infectious diseases in my country after the 1990s, the incidence of intestinal infectious diseases has always been among the best. Vibrio cholerae, Salmonella, Shigella, Shigella, Vibrio parahaemolyticus, Bacillus cereus, Campylobacter, Clostridium perfringens, Sakazakii, Escherichia coli, Staphylococcus aureus, monocytes Listeria monocytogenes, Escherichia coli O157 and Norovirus I, Norovirus II, Rotavirus A, Rotavirus B, Rotavirus C and other eighteen pathogens are important pathogens causing foodborne infections , often causing large-scale outbreaks i...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/14C12Q1/10C12Q1/04
CPCC12Q1/6837C12Q2531/113C12Q2565/501Y02A50/30
Inventor 田桢干张子龙张晓航李深伟王俐李平
Owner SHANGHAI ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF P R C
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