Kit and method for detecting vibrio parahaemolyticus and integron in food

A technology for hemolytic vibrio and integrons, which is applied in the field of probes, can solve the problems of unreported detection of vibrio parahaemolyticus and integrons, and achieve good industrialization prospects, good repeatability, and convenient use

Inactive Publication Date: 2016-11-09
INSPECTION AND QUARANTINE TECHNOLOGY CENTER ZHONGSHAN ENTRY EXIT INSPECTION AND QUARANTINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] However, there are few reports on the analysis of copy number based on the droplet digital PCR platform, let alone the detection of Vibrio parahaemolyticus and integrons

Method used

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  • Kit and method for detecting vibrio parahaemolyticus and integron in food
  • Kit and method for detecting vibrio parahaemolyticus and integron in food
  • Kit and method for detecting vibrio parahaemolyticus and integron in food

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Experimental program
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Effect test

Embodiment 1

[0051] The present invention detects the primers of Vibrio parahaemolyticus and the integron (int1) in food, and the sequences of the primers are respectively:

[0052] Upstream primer VPF: GCCACACTGGAACTGAGACA

[0053] Downstream primer VPR: GGAGTTAGCCGGTGCTTCTT

[0054] Probe VPP: TGCATTATTTGACGTTAGCGAC

[0055] Upstream primer int1F: CATCGTCGTAGAGACGTCGG

[0056] Downstream primer Int1R: AGAACAAGCAGGCATCACGA

[0057] Probe IntlP: AGGGTGTGCGGTGTGGCGGGC.

Embodiment 2

[0059] The present invention is a kit for detecting Vibrio parahaemolyticus and integron int1 in food, wherein the 20 μL reaction system includes the following components:

[0060] 2×ddPCR Super Mix 10.0 μL, Vibrio parahaemolyticus and int1 forward and reverse primers 1.0 μL each, probe 1.0 μL each, DNA template 4.0 μL.

[0061] Wherein the primer sequences are as follows:

[0062] Upstream primer VPF: GCCACACTGGAACTGAGACA

[0063] Downstream primer VPR: GGAGTTAGCCGGTGCTTCTT

[0064] Probe VPP: TGCATTATTTGACGTTAGCGAC

[0065] Upstream primer int1F: CATCGTCGTAGAGACGTCGG

[0066] Downstream primer Int1R: AGAACAAGCAGGCATCACGA

[0067] Probe IntlP: AGGGTGTGCGGTGTGGCGGGC.

Embodiment 3

[0069] The method for detecting vibrio parahaemolyticus and integron int1 in food of the present invention comprises the following steps:

[0070] A. Extract sample DNA;

[0071] Add each reaction component to 20.0 μl reaction system, then add 70.0 μl mineral oil, mix well and transfer to a droplet generator to automatically generate droplets;

[0072] The 20 μL reaction system includes the following components:

[0073] 2×ddPCR Super Mix 10.0 μL, Vibrio parahaemolyticus and int1 forward and reverse primers 1.0 μL each, probe 1.0 μL each, DNA template 4.0 μL.

[0074] Wherein the primer sequences are as follows:

[0075] Upstream primer VPF: GCCACACTGGAACTGAGACA

[0076] Downstream primer VPR: GGAGTTAGCCGGTGCTTCTT

[0077] Probe VPP: TGCATTATTTGACGTTAGCGAC

[0078] Upstream primer int1F: CATCGTCGTAGAGACGTCGG

[0079] Downstream primer Int1R: AGAACAAGCAGGCATCACGA

[0080] Probe IntlP: AGGGTGTGCGGTGTGGCGGGC.

[0081] B. Carefully transfer all the generated micro-droplets...

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Abstract

The invention discloses a primer, a probe, a kit and a method for detecting vibrio parahaemolyticus and integron in food. The sequence of the primer is: forward primer VPF: GCCACACTGGAACTGAGACA, and reverse primer VPR: GGAGTTAGCCGGTGCTTCTT; the sequence of the integron primer is: forward primer int1F: CATCGTCGTAGAGACGTCGG, and reverse primer Int1R: AGAACAAGCAGGCATCACGA. The primer, the probe and the other components in the kit provided by the invention are reasonable in coordination, convenient in use and quick and accurate in detection. The method provided by the invention has simple and quick operation and low cost. The detection result is accurate.

Description

technical field [0001] The present invention relates to a primer for detecting Vibrio parahaemolyticus and integrons in food, and a probe used in conjunction with the primer; the present invention also relates to a double droplet digital PCR detection reagent comprising the above primer and probe The present invention also relates to a method for detecting Vibrio parahaemolyticus and integron int1 in food by using the kit. Background technique [0002] Vibrio Parahemolyticus is a gram-negative bacillus with arc-shaped, rod-shaped, filamentous and other shapes, without dentition. Eating food containing this bacteria can cause food poisoning, also known as halophilic bacteria food poisoning. Vibrio parahaemolyticus is a marine bacterium mainly derived from seafood such as fish, shrimp, crab, shellfish and seaweed. Vibrio parahaemolyticus is mainly transmitted through food, and it often needs to multiply to a certain number to cause disease. It usually causes infection and di...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/63
CPCC12Q1/689C12Q1/6851C12Q2600/16C12Q2531/113C12Q2563/159C12Q2537/143
Inventor 蔡先全吴冰王勇邱德义李蓉邱霞张杰豪萧崎倩
Owner INSPECTION AND QUARANTINE TECHNOLOGY CENTER ZHONGSHAN ENTRY EXIT INSPECTION AND QUARANTINE
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