Campylobacter jejuni rapid quantitative detection kit and using method

Abstract

The invention relates to the technology of biological detection, and aims to provide a campylobacter jejuni rapid quantitative detection kit. The kit comprises a plurality of freezing tubes, wherein the freezing tubes A are filled with freeze-dried powder, the freeze-dried powder comprises the following ingredients: a probe-method fluorescent quantitative PCR pre-mixed liquid reagent as well as two primers and a probe which are used for fluorescent quantitative detection; the five freezing tubes B are respectively filled with sterile double distilled water; the five freezing tubes C are respectively filled with TZ lysate; the total number of the freezing tubes D, E, F, G, H, I and J is seven, the freezing tubes D, E, F, G, H, I and J are respectively filled with campylobacter jejuni standard substance DNA freeze-dried powder with different concentrations, and the concentration of the campylobacter jejuni is decreased progressively by ten times. The campylobacter jejuni rapid quantitative detection kit is realized on the basis of design of primer probes on campylobacter jejuni mapA genes and optimization of detection conditions. By a campylobacter jejuni rapid quantitative detection method, the number of campylobacter jejuni in a quantitative sample can be detected within a short time, and the minimum lower limit of detection can reach 102 CFU/mL; the campylobacter jejuni rapid quantitative detection method has good specificity, sensitivity and accuracy, is convenient and speedy, and is easy to operate.

Description

technical field [0001] The invention belongs to biological detection technology, in particular to rapid and quantitative detection of Campylobacter jejuni in food. The aim is to establish a rapid detection technology based on fluorescent quantitative PCR for Campylobacter jejuni, that is, a rapid quantitative detection kit for Campylobacter jejuni and its application method. Background technique [0002] Campylobacter jejuni belongs to the genus Campylobacter, a Gram-negative, microaerophilic, spiral-shaped bacterium that mainly causes acute enteritis in humans, accompanied by reactive arthritis, hepatitis, Reiter's disease and Guillain-Barré syndrome diseases such as immunocompromised diseases. In 1980, the World Health Organization (WHO) listed campylobacter enteritis as one of the most common infectious diseases. In 2003, my country's National Foodborne Disease Surveillance Network will also list Campylobacter as a monitoring object. [0003] In many countries, the col...

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Problems solved by technology

[0004] The traditional culture method is commonly used to detect Campylobacter. The traditional method takes a long time, and it usually takes 5 to 6 days to get the test result.

Campylobacter jejuni can survive in the environment for several weeks in a "viable non-culturable" state, and traditional detection methods cannot effectively detect these bacteria

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