RDA (Recombinase-dependent amplification) method and kit for quickly detecting Coxsachie virus 16 and enterovirus 71

Abstract

The invention discloses an RDA (Recombinase-dependent amplification) method and a kit for quickly detecting a Coxsachie virus 16 and an enterovirus 71. The kit comprises a specific primer pair and anRDA fluorescence labeling probe to realize a purpose of safely, specifically, sensitively and conveniently detecting the Coxsachie virus 16 (CA 16) and an enterovirus 71 (EV 71) so as to make up for deficiencies in an existing traditional detection technology. The kit provided by the method can omit a nucleic acid extraction step, and the detection of the CA 16 and the EV 71 can be realized in 20min under a constant temperature condition of 37-42 DEG C, specificity is 100%, and the kit is very suitable for field quick detection. Compared with a common PCR (Polymerase Chain Reaction) method, the RDA fluorescence method carries out reaction at a constant temperature, does not require temperature change, does not require a complex instrument and is short in reaction time. Therefore, the method and the kit thereof have the characteristics of being simple and quick in operation, good in specificity, high in sensitivity, low in cost and the like, provide an effective technical means for thefield quick detection and screening of the CA 16 and the EV 71, and have a wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of molecular biology. More specifically, it relates to a pair of primers, probes and related kits for detecting Coxsackievirus A16 and Enterovirus 71 nucleic acids based on RDA fluorescence detection technology. Background technique [0002] Hand-foot-and-mouth disease (Hand-foot-and-mouth disease, HFMD) is an infectious disease caused by enteroviruses. There are more than 20 enteroviruses (types) that cause HFMD, among which Coxsackievirus A16 type ( Coxsachie virus 16, CA16) and enterovirus 71 (Enterovirus 71, EV71) are the most common. Hand, foot and mouth disease is a common infectious disease in infants and young children. It mostly occurs in children under 5 years old. heal. HFMD virus can be transmitted through direct contact with nasopharyngeal secretions, saliva, herpes fluid or feces of an infected person. Enterovirus infection, the incubation period is 2 to 10 days, with an average of 3 to 5 d...

AI Technical Summary

Problems solved by technology

Among them, the virus isolation detection method has a long cycle and is not suitable for clinical detection, and the reliability of the serum immune detection method is poor. PCR detection methods such as conventional PCR, RT-PCR, real-time fluorescent quantitative PCR, etc., play an important role in the detection of pathogenic microorganisms, but their general It requires instruments and laboratories capable of precise temperature control, and has high operating requirements for testing personnel. It is not suitable for on-site testing of samples, is not conducive to grassroots promotion, and cannot meet the simple, fast and accurate testing requirements for disease prevention and control.

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