Primers for screening and parting enterovirus viruses and detection method

Abstract

The invention discloses primers for screening and parting enterovirus viruses and a detection method. The sequences of the primers are the CODEHOP outer primer sequence, the CODEHOP inner primer sequence and the primer sequences of 20 viruses. The detection method includes the steps of extraction of RNA; one-step RT-PCR; nest type PCR; 1.5% agarose gel electrophoresis to judge the positivity and parting result of enteroviruses. A pair of enterovirus consistent-degenerate primers is designed, a pair of short-fragment consistent-degenerate primers and parting primers of 20 different enteroviruses are designed inside amplified fragments of the primers, and in combination with the nest type RT-PCR technology, a degenerate primer nest type RT-PCR method capable of being used for screening and parting different genotype viruses of the enteroviruses is established. The advantages of being high in sensitivity and specificity and suitable for monitoring and variety identifying of enteroviruseses of low-virus-content samples are achieved.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to a primer and a detection method for screening and typing Enteroviruses. Background technique [0002] In recent years, outbreaks of human infectious diseases caused by enterovirus infection have often been reported, and have become an important public health problem; Infect humans, including poliovirus, coxsackievirus, echovirus and some new enteroviruses and hepatitis A virus. For example, in 1998, hand, foot and mouth disease and herpetic pharyngitis infected by enterovirus type 71 (EV71) broke out in Taiwan. The number of infected people exceeded 100,000. Edema and hemorrhage, acute flaccid paralysis and myocarditis died. At present, HEV accounts for about 80% to 92% of the pathogenic factors of aseptic meningitis. In recent years, some studies have shown that diabetes is also related to enteroviruses. Enteroviruses carried by patients and infected ...

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a primer and detection method for screening and typing of enteroviruses, aiming to solve the problem of high cost, time-consuming, cumbersome operation and easy contamination of existing enterovirus detection technology, which is difficult for unknown Types of viruses are easy to miss and cannot be used for virus typing, and the sensitivity of samples is not enough to easily cause false negatives

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