Double LAMP (loop-mediated isothermal amplification) detection primer and kit for Vibrio parahaemolyticus and Vibrio cholera in aquatic products

A hemolytic vibrio and detection kit technology, which is applied in the direction of recombinant DNA technology, microbial measurement/inspection, and resistance to vector-borne diseases, etc., can solve the problems of increased non-specific amplification reactions, decreased sensitivity, and high detection costs. Achieve the effect of less non-specific amplification reaction, reduce the cost of inspection, and improve the detection speed

Inactive Publication Date: 2016-04-13
舟山出入境检验检疫局综合技术服务中心
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  • Abstract
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This technology has been preliminarily studied in the detection of food-borne pathogens, but all of them use a single detection, which takes a long time and costs high
The dual LAMP system for simultaneous detection of Vibrio parahaemolyticus and Vibrio cholerae has not been reported
If a multiple LAMP system is used, due to the existence of multiple pairs of primers, the interference between primers and the mismatch between primers and templates will cause problems such as decreased sensitivity and increased non-specific amplification reactions.

Method used

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  • Double LAMP (loop-mediated isothermal amplification) detection primer and kit for Vibrio parahaemolyticus and Vibrio cholera in aquatic products
  • Double LAMP (loop-mediated isothermal amplification) detection primer and kit for Vibrio parahaemolyticus and Vibrio cholera in aquatic products
  • Double LAMP (loop-mediated isothermal amplification) detection primer and kit for Vibrio parahaemolyticus and Vibrio cholera in aquatic products

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Experimental program
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Embodiment

[0045] A double LAMP detection kit for Vibrio parahaemolyticus and Vibrio cholerae in aquatic products, comprising a double LAMP amplification reaction liquid, the double LAMP amplification reaction liquid is mixed with the following components in a total volume of 25 μL:

[0046] 12.5 μL of reaction buffer, 1 μL of internal primer ompw-FIP with a concentration of 20 μmol / L, 1 μL of internal primer ompw-BIP with a concentration of 20 μmol / L, 1 μL of internal primer toxR-FIP with a concentration of 20 μmol / L, 1 μL of internal primer toxR-BIP with a concentration of 20 μmol / L, The outer primer ompw-F30.125 μL with a concentration of 20 μmol / L, the outer primer ompw-B30.125 μL with a concentration of 20 μmol / L, the outer primer toxR-F30.125 μL with a concentration of 20 μmol / L, and the outer primer toxR-B30 with a concentration of 20 μmol / L. 125 μL, loop primer ompw-LF at a concentration of 20 μmol / L 0.5 μL, loop primer ompw-LB at a concentration of 20 μmol / L 0.5 μL, loop primer t...

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Abstract

The invention discloses a double LAMP (loop-mediated isothermal amplification) detection primer and kit for Vibrio parahaemolyticus and Vibrio cholera in aquatic products, comprising three pairs of Vibrio cholera LAMP primers and three pairs of Vibrio parahaemolyticus LAMP primers. Inter-primer interference is low, nonspecific amplification reactions are few, detection sensitivity and specificity are guaranteed, detection speed is increased, and detection cost is lowered.

Description

technical field [0001] The invention relates to the technical field of biological detection of aquatic products, in particular to a dual LAMP detection primer and a kit for Vibrio parahaemolyticus and Vibrio cholerae in aquatic products. Background technique [0002] With the improvement of people's living standards, people pay more and more attention to food safety, which has become a major worldwide public problem, and foodborne diseases caused by pathogenic microorganisms are one of the most important factors affecting food safety. Vibrio parahaemolyticus and Vibrio cholerae are recognized worldwide as two important intestinal pathogens. The former widely exists in the marine environment and is one of the main pathogenic bacteria threatening the mariculture industry. It can infect fish, shrimp and shellfish, etc., cause shrimp red body disease, fish skin ulcers and other diseases, and cause major economic losses to the aquaculture industry. . Humans can cause food poiso...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12N15/11
CPCC12Q1/6844C12Q1/689C12Q2600/16C12Q2537/143C12Q2531/119Y02A50/30
Inventor 相兴伟胡兴娟邵宏宏周秀锦周向阳廖妙飞洪瑶
Owner 舟山出入境检验检疫局综合技术服务中心
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