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Method for detecting drug-resistant mutation sites of campylobacter jejuni carbostyril antibiotics

A technology of campylobacter jejuni and mutant type, applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of high cost, high culture conditions, and time-consuming, and achieve good specificity and sensitivity, The effect of accurate medication guidance and simplification of complex procedures

Active Publication Date: 2014-04-02
CHINA AGRI UNIV
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Problems solved by technology

Traditional genotype mutation detection methods mainly include polymerase chain reaction-linear probe method, polymerase chain reaction-restriction fragment length polymorphism analysis method, mismatch amplification mutation polymerase chain reaction method and fluorescence quantitative polymerization Enzyme chain reaction and other methods, the above-mentioned genotype mutation detection methods based on polymerase chain reaction are relatively expensive, and are not suitable for detecting a large number of samples, and they are only qualitative tests, which cannot be used for point mutations. qualitative test
The traditional drug susceptibility test requires the culture of Campylobacter jejuni, which requires high culture conditions and takes a lot of time, which does not meet the needs of emergency prevention and control of acute infectious diseases

Method used

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  • Method for detecting drug-resistant mutation sites of campylobacter jejuni carbostyril antibiotics
  • Method for detecting drug-resistant mutation sites of campylobacter jejuni carbostyril antibiotics
  • Method for detecting drug-resistant mutation sites of campylobacter jejuni carbostyril antibiotics

Examples

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Embodiment 1

[0027] Example 1. Using high-resolution melting curve analysis to assist in the detection or detection of whether the 257th position of the gyrA gene coding sequence of Campylobacter jejuni is C or T

[0028] In this example, Roche LightCycler480 real-time fluorescent quantitative PCR system (the system includes Roche 480 fluorescent quantitative PCR instrument and Roche’s Gene Scanning module) was used for high-resolution melting curve analysis to establish auxiliary detection or detection of Campylobacter jejuni gyrA The method of whether the 257th position of the gene coding sequence is C or T. details as follows:

[0029] 1. Design of PCR amplification primers in high resolution melting curve analysis

[0030] jejuni NCTC11168 strain, Campylobacter jejuni ZP11, Campylobacter jejuni standard quality control strain ATCC33560, quinolone-resistant Campylobacter jejuni HT8, Campylobacter jejuni LY10, Campylobacter jejuni LY17, Campylobacter jejuni SX4, Campylobacter jejuni Co...

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Abstract

The invention discloses a method for detecting drug-resistant mutation sites of campylobacter jejuni carbostyril antibiotics. The method comprises the following steps: by taking campylobacter jejuni of which the 257th site of a gyrA gene coding sequence is C as a wild type standard strain, taking campylobacter jejuni of which the 257th site of the gyrA gene coding sequence is T as a mutant type standard strain, respectively and simultaneously performing high-resolution melting curve analysis with campylobacter jejuni to be detected; determining whether the 257th site of the gyrA gene coding sequence of the campylobacter jejuni to be detected is C or T according to the comparison result of the high-resolution melting curve; if the high-resolution melting curve of the campylobacter jejuni to be detected is consistent with that of the wild type standard strain, selecting the campylobacter jejuni to be detected as a strain of which the 257th site of the gyrA gene coding sequence is C, and if the high-resolution melting curve of the campylobacter jejuni to be detected is consistent with that of the mutant type standard strain, selecting the campylobacter jejuni to be detected as a strain of which the 257th site of the gyrA gene coding sequence is T.

Description

technical field [0001] The invention relates to a method for detecting quinolone antibiotic resistance mutation sites of Campylobacter jejuni. Background technique [0002] Campylobacter jejuni (Campylobacter jejuni), also known as Campylobacter jejuni, is one of the most important food-borne pathogens that cause acute gastroenteritis in humans. Caused by Campylobacter coli. Campylobacter jejuni can be transmitted to humans through the food chain, and contaminated poultry meat, milk and drinking water are the main routes of transmission. Quinolones are the most commonly used drugs in the treatment of enteritis caused by Campylobacter jejuni, and they are also the first-line treatment drugs for Campylobacter infection in clinical practice. The emergence of drug-resistant Campylobacter jejuni leads to clinically protracted course of Campylobacter jejuni disease and treatment failure, which poses a great threat to human health. At the same time, drug-resistant strains also le...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/04C12N15/11C12R1/01
CPCC12Q1/6858C12Q2531/113C12Q2527/107C12Q2545/113
Inventor 吴聪明汪洋吴辰斌沈建忠
Owner CHINA AGRI UNIV
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