Primer pair combination and kit for detecting and identifying human tissue echinococcosis pathogen

Abstract

The invention discloses a multiple PCR method for simultaneously detecting and identifying three human tissue echinococcosis pathogens, including echinococcus granulosus G1-G3, echinococcus multilocularis and echinococcus granulosus G6-G10. According to the invention, 3 pairs of specific primers are designed for the mitochondrial genes of 3 pathogens; the specific primers are added into a same PCR reaction system; the specific primers are complementarily combined with the target genes of the corresponding parasite species; through amplified reaction, different parasite species can generate target stripes in different sizes; gel electrophoresis is adopted for separating and detecting. The three pairs of primers according to the invention are free from mutual interference and have no cross reaction with 8 tapeworms in close genetic relationship or do not interfere with the result judgment. The method is high in specificity and sensitivity, can effectively and accurately realize the synchronous detection for 3 pathogens, can greatly save the detection time and cost, can effectively increase the working efficiency and can be applied to the research on the parting identification for human echinococcosis pathogens and echinococcosis molecular epidemiology.

Description

technical field [0001] The invention belongs to the field of disease diagnosis, and in particular relates to a primer pair combination and a kit for detecting and identifying various human tissue echinococcosis pathogens. Background technique [0002] Hydatidosis, commonly known as echinococcosis, is a zoonotic parasitic disease caused by the larvae of Echinococcus tapeworms parasitic in the human body. Echinococcus genus found in the world in the past is divided into 6 species, namely Echinococcus granulosus (Echinococcus granulosus), Echinococcus multilocularis (Echinococcus multilocularis), Echinococcus vogeli (Echinococcus vogeli), Echinococcus oligonodes (Echinococcus oligarthus), Echinococcus felids and Echinococcus shiquicus (Echinococcus shiquicus), among which E. granulosus (E. granulosus) is divided into 10 genotypes according to genotype, that is, Echinococcus granulosus G1-G10 type. With the application and development of molecular biology techniques in the stud...

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Problems solved by technology

However, on the one hand, the existing multiplex PCR methods are not completely aimed at the three human echinococcosis pathogens found in my country (E. granulosus sensu stricto, E. Echinococcus G6–G10 type E. canadensis), which cannot meet the needs of detection and identification of human echinococcosis pathogens; Many pairs of primers are easy to cross-react with closely related pathogens, resulting in false positive results, and complementary binding between primer pairs is easy to occur, and interference reactions are easy to occur. Therefore, the existing multiplex PCR method for echinococcosis has specificity. Poor performance, unreliable methods, etc.

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