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Waters frequent food born pathogenic hacteria multiple PCR rapid detecting kit and its detecting method

A food-borne pathogenic bacteria and detection kit technology is applied in the field of Pseudomonas aeruginosa, Enterohemorrhagic Escherichia coli 0157 and Vibrio parahaemolyticus, and can solve the problems of many drug reagents, time-consuming and laborious, and high detection cost, Achieve the effect of short cycle, strong operability and low detection cost

Inactive Publication Date: 2006-08-09
GUANGDONG INST OF MICROORGANISM +1
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AI Technical Summary

Problems solved by technology

However, the existing PCR method only targets one pathogenic bacteria for each amplification. If there is a water sample to detect whether it contains one or more of the above pathogenic bacteria, it is necessary to perform multiple PCR reactions respectively. Therefore, it is time-consuming and laborious, consumes a lot of pharmaceutical reagents, and the detection cost is high

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  • Waters frequent food born pathogenic hacteria multiple PCR rapid detecting kit and its detecting method
  • Waters frequent food born pathogenic hacteria multiple PCR rapid detecting kit and its detecting method
  • Waters frequent food born pathogenic hacteria multiple PCR rapid detecting kit and its detecting method

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Embodiment

[0070] Example: multiplex PCR rapid detection kit for common foodborne pathogenic bacteria in water bodies

[0071] 1. Water sample collection and pretreatment

[0072] Collect 500ml of water samples in a sterile sampling bottle, centrifuge at 7000r / min for 20min, wash the precipitate with sterile distilled water three times, and then prepare DNA.

[0073] 2. DNA extraction

[0074] (1) Add 9.5mL TE to suspend the precipitate, add 0.5ml 10% SDS, 50μL 20mg / ml (or 1mg dry powder) proteinase K, mix well, and incubate at 37°C for 1 hour;

[0075] (2) Add 1.5ml 5mol / L NaCl and mix well;

[0076] (3) Add 1.5ml CTAB / NaCL solution, mix well, and keep warm at 65°C for 20 minutes; (4) Extract with an equal volume of phenol:chloroform:isoamyl alcohol (25:24:1), centrifuge at 5000r / min for 10 minutes, Transfer the supernatant to a clean centrifuge tube. ;

[0077] (5) Extract with an equal volume of chloroform:isoamyl alcohol (24:1), and transfer the supernatant to a clean tube;

[...

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Abstract

This invention relates to a multiple PCR quick test reagent box of food- pathogens often seen in water including 10xPCR reaction buffer solution, MgCl2, dNTP, a primer and Hot star Taq enzymes. The test method includes: picking up due test sample templates, adding 10xPCR buffer solution, MgCl2, dNTP, primers, the Hot star Taq enzymes, the due test sample and water in a PCR thin-wall tube to be mixed and put on the PCR instrument to be enlarged in an electrophoresis device, finally the result is analyzed and judged.

Description

[Affiliated technical field] [0001] The present invention relates to the detection method of common food-borne pathogenic bacteria and the synchronous detection multiplex PCR technology of multiple pathogenic bacteria genes in the water body, particularly relates to a kind of can be used for simultaneous detection Salmonella (Salmonella sp.), Shigella (Shigella sp. .), Pseudomonas aeruginosa, enterohaemorrhagic Escherichia coli 0157 (Eterohaemorrhagic 0157) and vibrio parahaemolyticus (Vibriorahaemolyticus) and other five kinds of PCR detection kits A technical method for food-borne pathogenic bacteria belongs to the field of biotechnology. [Background technique] [0002] Foodborne pathogens refer to a large group of bacteria that use food as a carrier to cause human diseases. In recent years, global food safety incidents have occurred frequently, such as the "Listeria incident" in the United States and the "Escherichia coli 0157 epidemic incident" in Japan, which have brou...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
Inventor 吴清平范宏英张菊梅郭伟鹏
Owner GUANGDONG INST OF MICROORGANISM
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