Multiplex fluorescence PCR detection kit for hand-foot-and-mouth disease viruses and application thereof

Abstract

The invention relates to a multiplex fluorescence PCR detection kit for hand-foot-and-mouth disease viruses and an application thereof. The multiplex fluorescence PCR detection kit comprises primer pairs used for amplifying common hand-foot-and-mouth disease pathogens as well as a primer and a probe for an internal standard gene used for quality control, sequences of the primer and the probe are shown in SEQ ID NO.1-SEQ ID NO.12. The multiplex fluorescence PCR detection kit provided by the invention has the advantages that three common pathogens resulting in intestinal system infection can bedetected at the same time during reaction at a time, detection time is shortened, accuracy is high, and a powerful detection tool is provided for clinic and detection laboratories; the multiplex fluorescence PCR detection kit provided by the invention has good specificity on a universal type primer and a probe which are used for an enterovirus type 71, a Coxsackie virus A16 and enteroviruses, andthe problem that different primers / probes are mutually cross-linked is avoided; besides, an internal standard is designed in the kit, an internal standard template is a human house-keeping gene and participates in sample extraction and PCR amplification, the whole reaction process can be effectively monitored, and false negative is prevented; and meanwhile, inhibition of some inhibitors on PCR reaction can be effectively evaded.

Description

technical field [0001] The invention relates to a kit, in particular to a multiple fluorescent PCR detection kit for hand, foot and mouth disease virus, and its application. Background technique [0002] Hand, foot and mouth disease (HFMD) is one of the common infectious diseases caused by a variety of enteroviruses. The disease has the characteristics of rapid spread and high prevalence, and can cause large-scale epidemics in a short period of time. It is more common in children under 5 years old, and the incidence rate is highest in the age group younger than 3 years old. From April to September every year, infants and young children are more likely to be infected with hand, foot and mouth disease, and the total number of patients generally reaches its peak in May to July. The main modes of transmission are: close contact with people, through contact with virus-contaminated toys, towels, milk utensils, underwear, bedding, etc.; the virus in the patient's throat secretions...

AI Technical Summary

Problems solved by technology

However, traditional fluorescent real-time PCR can only amplify the nucleic acid of one pathogen at a time. It takes multiple attempts to accurately identify the causative pathogen, which consumes a lot of time and energy, and it cannot effectively distinguish infections caused by multiple pathogens. In addition, it can only detect EV71, CA16 and other enteroviruses with a single index in different reaction tubes, and cannot detect all viruses at one time. Not only the detection efficiency is low, but also the detection cost is high

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