Primer, kit and method for detecting chlamydia

Abstract

The invention discloses an animal chlamydia detecting method based on an RPA (Recombinant Polymerase Amplification) technology. According to the method, recombinase and single-strand binding protein cooperate with each other at normal temperature to realize specific binding of a primer and a template, and a new DNA chain is generated by extending the primer under the action of DNA polymerase. Theanimal chlamydia detecting method is characterized in that a set of primer and probe combination used for RPA detection is designed and screened according to a chlamydia gene sequence envB, and an animal chlamydia specificity fluorescence RPA detecting method is established. The chlamydia specificity fluorescence RPA detecting method disclosed by the invention is easy, convenient and quick to operate and high in specificity, and is suitable for on-site quick detecting.

Description

technical field [0001] The present invention relates to a primer and a related probe for detecting chlamydia in animals, and a detection kit, specifically a specific fluorescent recombinase polymerase amplification technology (RPA) detection primer and a test kit for detecting chlamydia in animals, The invention also relates to a method for detecting animal chlamydiosis using the primer and probe or the kit of the invention for non-disease diagnosis. Background technique [0002] Chlamydia (Chlamydia) is a strictly intracellular parasitic Gram-negative bacterium with a diameter of 0.2-0.5 μm, which is between Rickettsia and viruses, and can pass through a 0.45 μm filter membrane. It can only grow in host cells and cannot Most bacteria grow on agar plates as well. The pathogen of chlamydia has a wide range of pathogenicity. In recent years, chlamydia has become more prevalent in my country, especially in dairy cows, yaks, sheep, goats, pigs, etc., which has seriously hindere...

AI Technical Summary

Problems solved by technology

Among them, the two methods of indirect hemagglutination diagnostic test and complement fixation test have the characteristics of low sensitivity, poor specificity, strong subjectivity in judgment of results, and unsuitability for large-scale detection, which limits their wide application in clinical practice.

ELISA diagnostic method is mainly aimed at lipopolysaccharide antigens, but some epitopes of Chlamydia lipopolysaccharide antigens are similar to those of other Gram-negative bacteria, and cross-reactions and false positives are prone to occur

For example, the prior art CN201110367644.0 discloses an ELISA kit for detecting antibodies to Chlamydia abortifaciens, which uses recombinant Chlamydia main outer membrane protein rMOMP as the coating antigen, and in the prior art uses recombinant pleomorphic outer membrane protein POMP The indirect ELISA antibody detection method for porcine abortion chlamydial disease established for the coated antigen, but these existing technologies are all ELISA antibody indirect detection methods or kits developed for a specific animal, which have a narrow scope of application and are not universal. It cannot be used for the detection and diagnosis of chlamydia in various animals, the cost of use is high, and it is difficult to popularize

Micro-volume indirect immunofluorescence test was once recommended as the most reliable serological method for the diagnosis of Chlamydia pneumoniae, but this diagnostic method requires high technical requirements and takes a long time, so it has not become a common detection item for Chlamydia pneumoniae

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