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Detection kit for vibrio alginnolyficus

A detection kit and Vibrio alginolyticus technology, which is applied in the field of pathogenic bacteria diagnosis, can solve the problems of time-consuming and laborious, high requirements for operators, and cumbersome methods

Inactive Publication Date: 2014-03-05
胡成进
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the traditional methods for detecting marine pathogens mainly include pre-enrichment, selective enrichment, selective culture, biochemical identification, and serological identification. It takes 5 to 6 days to issue a report. The method is cumbersome, time-consuming and laborious.
The PCR method, although fast, sensitive, and specific, has high requirements for operators and requires special instruments, and is also not suitable for on-site detection.

Method used

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  • Detection kit for vibrio alginnolyficus
  • Detection kit for vibrio alginnolyficus
  • Detection kit for vibrio alginnolyficus

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Experimental program
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Embodiment 1

[0050] 1. Design and synthesis of primers

[0051] The homology analysis and comparison of the known sequences of the Vibrio alginolyticus genome in Genebank finally determined that the gyrB (EF542801.1) gene was a candidate gene for the diagnosis of Vibrio alginolyticus,

[0052] Table 4-1 Candidate genes for LAMP amplification of Vibrio alginolyticus

[0053] strain gene sequence number gene name Location fragment length Vibrio alginolyticus EF542801.1 gyrB 670~1019 349bp

[0054] Primers were designed according to the candidate genes, and the designed primers were synthesized by Shanghai Sangon Biotechnology Co., Ltd.;

[0055] 2. Reagent preparation

[0056] Add deionized water to the dry primer powder to make a 100uM preservation solution, and dilute the primer preservation solution ten times for use in experiments. Add 20ml of deionized water to Chelex100 to make a 5% Chelex100 solution;

[0057] 3. Sample processing

[0058] ...

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Abstract

The invention relates to a detection kit for vibrio alginnolyficus. The detection kit specifically comprises a primer and an LAMP (Loop-mediated isothermal Amplification) actual reaction liquid. The kit is high in sensitivity, strong in specificity, low in cost and simple and convenient to operate.

Description

technical field [0001] The invention belongs to the technical field of pathogenic bacteria diagnosis, and in particular relates to a rapid detection kit and a detection method for detecting Vibrio alginolyticus using LAMP technology. Background technique [0002] Vibrio alginolyticus is a Gram-negative opportunistic pathogen that usually infects humans through raw seafood consumption or wound contact with contaminated seawater or seafood. Clinical symptoms mainly manifest as diseases such as wound infection, gastroenteritis, otitis media, endophthalmitis, food poisoning and sepsis. At present, the traditional methods for detecting marine pathogens mainly include pre-enrichment, selective enrichment, selective culture, biochemical identification, and serological identification. It takes 5 to 6 days to produce a report, which is cumbersome, time-consuming and laborious. The PCR method, although fast, sensitive, and specific, has high requirements for operators and requires sp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/04C12Q1/6844C12Q2531/119Y02A50/30
Inventor 胡成进刘晓斐汤潜
Owner 胡成进