Primer set for rapid detection of gyra gene mutation in Campylobacter jejuni and its application

Abstract

The invention discloses a primer group for quickly detecting the gyrA gene mutation site of campylobacter jejuni and an application thereof. The primer group consists of a specific primer pair and a sequencing primer, wherein the specific primer pair consists of a single-strand DNA molecule A shown by the sequence 1 and a single-strand DNA molecule B shown by the sequence 2, and the sequencing primer is a single-strand DNA molecule C shown by the sequence 3. By adopting the campylobacter jejuni to be detected as a template and adopting the specific primer pair, a gyrA gene segment including a 257 mutation site is obtained through a PCR (polymerase chain reaction), and a DNA single strand can be obtained through single-strand separation and purification of PCR products and is directly put into a pyrosequencing apparatus for sequencing. By adopting the primer group disclosed by the invention, the C257T mutation of campylobacter jejuni can be identified so as to determine whether the campylobacter jejuni to be detected tolerates quinolone antibiotics at a high level. A powerful technical means is provided for the detection of anti-quinolone campylobacter jejuni as well as molecular epidemiological monitoring.

Description

technical field [0001] The invention relates to a primer set for rapidly detecting the gyrA gene mutation site of Campylobacter jejuni and its application. Background technique [0002] Campylobacter jejuni, also known as Campylobacter jejuni, is one of the most important food-borne pathogens that cause acute gastroenteritis in humans. More than 95% of clinical Campylobacter enteritis is caused by Campylobacter jejuni and 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 thr...

AI Technical Summary

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

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