Campylobacter jejuni fluorescence quantitative PCR detection kit and detection method thereof

Abstract

The invention provides a campylobacter jejuni fluorescence quantitative PCR detection method and a detection kit thereof. The method can rapidly and accurately detect the campylobacter jejuni and reduces the false positive. The kit comprises the following reagents: PCR MIX reaction liquid, 5*FQ PCR buffer, a primer mixture with the concentration of 10 mu M, a TaqMan probe mixture with the concentration of 10 mu M, and dNTPs and ddH2O with the concentration of 10 mM, wherein the primer mixture comprises a 16S rRNA upstream primer F, a 16S rRNA downstream primer R, and an upstream primer hipO-F and a downstream primer hipO-R; and the TaqMan probe mixture comprises: (1) 16S rRNA-P and hipO-P; (2) Taq enzyme; and (3) positive standard substances and negative standard substances. By introducing high-efficiency and strong-specificity amplification primers and fluorescent probes, the sensitivity and specificity of detecting the campylobacter jejuni are improved and missing detection and error detection are avoided.

Description

technical field [0001] The invention relates to a nucleic acid detection kit for Campylobacter jejuni and a detection method thereof, belonging to the field of biotechnology. Background technique [0002] Campylobacter jejuni (Campylobacter jejuni) is an important pathogenic bacterium that is widely prevalent in the world. It not only causes a variety of livestock and poultry diseases, but also causes food poisoning in humans. Studies have proved that people infected with Campylobacter jejuni, Most cases are directly or indirectly related to animal food. Among all kinds of bacterial food poisoning in various countries, food poisoning caused by Campylobacter jejuni is often in the forefront, so the World Health Organization has listed this disease as one of the most common foodborne diseases. [0003] Campylobacter jejuni can be found in many types of food, including raw meat, poultry, eggs, dairy products, fish, shrimp and other foods. Campylobacter jejuni is in a subletha...

AI Technical Summary

Problems solved by technology

Campylobacter jejuni is sublethal after being injured during food processing, and enters a "non-culturable state" in water, thus further increasing the possibility of contamination and the difficulty of detection

At present, the kit screening method for the detection of Campylobacter jejuni reported at home and abroad, although the operation steps are reduced and the detection target is narrowed down, there are still problems such as false positives and high cost.