Method for simultaneously detecting various intestinal tract viruses

Abstract

The invention discloses a method for simultaneously detecting various intestinal tract viruses, and belongs to the field of biologic detection. The method comprises the steps of respectively designinga pair of specific primers Fs and Rs and general primers Fg and Rg for target genes of intestinal tract virus general type, coxsackie virus A4 types, A6 types, A10 types and A16 types, and intestinaltract virus 71 types; firstly performing enrichment amplification through specific primers Fg+Fs and Rg+Rs with general primer labels; then performing exponential amplification with general primer labels Fg and Rg; and collecting fluorescence signals of exponential amplification products for detection. Through the adoption of the method disclosed by the invention, 6 kinds of intestinal tract virus pathogens of the intestinal tract virus general type, the coxsackie virus A4 types, A6 types, A10 types and A16 types, and the intestinal tract virus 71 types can be identified at a time. The methodis high in specificity and high in sensitivity, realization of early discovery, early diagnose and early treatment of hand-foot-mouth diseases can be facilitated, and besides, emergency disposal andpreventing and controlling capacity for hand-foot-mouth disease epidemic situations can be notably improved.

Description

technical field [0001] The invention relates to the field of biological detection, in particular to a method for simultaneously detecting multiple enteroviruses. Background technique [0002] Hand-foot-mouth disease (HFMD), also known as exanthematous stomatitis, is caused by enterovirus infection, and its clinical manifestations are mainly characterized by fever and rashes or herpes on the hands, feet, and oral cavity. An acute common infectious disease. It is more common in children, and infants and young children under 3 years old are more likely to get sick. HFMD is highly contagious, with complex transmission routes and rapid transmission, which can cause a pandemic in a short period of time. The pathogens causing HFMD are mainly types 2, 4, 5, 7, 9, 10, and 16 of the Coxsackie virus group A of the Enterovirus genus, and types 1, 2, 3, and 16 of the Coxsackie virus group B. Types 4 and 5; some ECHO-viruses, Enterovirus 71. The more common ones are Coxsackievirus A g...

AI Technical Summary

Problems solved by technology

However, it is difficult for traditional multiplex PCR technology to simultaneously amplify multiple target sequences with multiple pairs of primers, because each different target sequence has its own optimal amplification conditions, and it is difficult to unify them. Target sequences also have different amplification efficiencies, resulting in low concentration / titer pathogens not being amplified and not detected, resulting in false negative results

In addition, labeled primers tend to dimerize when used at high concentrations, causing high background in amplification results

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