Primer probe group and kit for detecting nucleic acid of schistosoma japonicum katsurada based on fluorescent RMA method and detection method of schistosoma japonicum katsurada

Abstract

The invention belongs to the technical field of biological detection, and particularly relates to a primer probe group, a kit and a detection method for detecting nucleic acid of schistosoma japonicum katsurada based on a fluorescent RMA method. The primer probe group comprises a primer pair for detecting schistosoma japonicum katsurada and a corresponding probe, a fluorescence reporter group marked on the detection probe of schistosoma japonicum katsurada is VIC, and a fluorescence quenching group is BHQ1; the kit comprises a detection tube containing an amplification reaction reagent, a buffer solution, magnesium acetate, a standard positive plasmid and sterile double distilled water, and the amplification reaction reagent is packaged in a single tube, is in a dry powder form, and comprises an RMA primer pair, a detection probe group, escherichia coli RecA protein, UvsY protein, single-chain binding protein GP32, Bst polymerase, exonuclease III, polyethylene oxide, trehalose, mannitol, ATP, dNTPs, creatine kinase and creatine phosphate.

Description

technical field [0001] The application belongs to the technical field of biological detection, and specifically relates to a primer probe set, a kit and a detection method for detecting Schistosoma japonicum nucleic acid based on the fluorescent RMA method. Background technique [0002] Schistosomiasis is a serious zoonotic parasitic disease that is widely distributed in Asia, Africa, South America and the Middle East. There are currently six types of schistosomes that parasitize humans: Schistosoma japonicum, Schistosoma mansoni, Schistosoma haematobium, Schistosoma intercalatum, Schistosoma mekong and Schistosoma malayi. The pathogen of schistosomiasis in my country is Schistosoma japonicum, which is mainly prevalent in the Yangtze River Basin and 13 provinces, municipalities and autonomous regions south of the Yangtze River in China. Schistosoma japonicum belongs to flatworm phylum, trematode class, dioecious order, Schistosomatae, Schistosoma genus, Schistosoma japonicu...

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Problems solved by technology

Traditional etiological diagnosis methods such as miracidium hatching method have a very low positive rate, missed detection and misdiagnosis are common; immunological diagnosis such as ring egg precipitation test, indirect hemagglutination test, enzyme-linked immunosorbent assay (ELISA), etc. Antibodies produced in the host body after infection have a long detection window and cannot distinguish between current infection and past infection; molecular biology methods are mainly fluorescent quantitative PCR and LAMP constant temperature amplification method, but the PCR method requires the use of complex instruments and well-equipped However, the LAMP method has high requirements on primers, and is particularly prone to aerosol pollution, causing false positives to affect the test results.

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