Qualitative detection method for simultaneously detecting three types of main hand-foot-and -mouth disease viruses in water environment

Abstract

The invention discloses a qualitative detection method for simultaneously detecting three types of main hand-foot-and-mouth disease viruses (EV71,CVA10,CVA16) in water environment, according to conservative property of nucleic acid in a section from a 5' noncoding region of RNA (Ribose Nucleic Acid) of the three types of main hand-foot-and-mouth disease viruses (EV71,CVA10,CVA16) to a viral protein capsid coding region VP2, and semi-nested PCR (Polymerase Chain Reaction) primers of the three types of main viruses of the hand-foot-and-mouth disease can be produced. A qualitative PCR detection method for the hand-foot-and-mouth disease viruses in the water environment by the semi-nested PCR primers can be built. The method has the advantages of good specificity and high sensitivity, and can be used for establishing a favorable base for follow-up genotyping.

Description

technical field [0001] The invention relates to a qualitative detection method for simultaneously detecting three main hand, foot and mouth disease viruses in water environment. The method designs general primers according to the conserved sequence of hand, foot and mouth disease virus RNA, and uses nested PCR to detect hand, foot and mouth disease virus. It is suitable for water Rapid detection of hand, foot and mouth disease virus in the environment. Background technique [0002] Reverse transcription-polymerase chain reaction (RT-PCR) technology uses RNA as a template to perform reverse transcription to obtain the first strand of complementary DNA (cDNA), and then use the cDNA strand as a template for PCR amplification reaction, thus A very small amount of RNA can be specifically amplified geometrically within a few hours. Semi-nested PCR (semi-nested PCR) technology is a variant of PCR technology that amplifies nucleic acid fragments by using a pair of half-PCR primers....

AI Technical Summary

Problems solved by technology

There are few existing PCR detection methods for hand, foot and mouth disease virus in the water environment, most of which are based on the qualitative or quantitative RT-PCR detection technology of enterovirus universal primers. Different types, there are disadvantages that the pathogen can not be quickly and accurately determined, the sensitivity is not high enough, and it is greatly affected by the interference of impurities

Although nested PCR technology has been applied to the detection of microbes in medical fields, it cannot meet the needs of water environment monitoring. Therefore, there is currently no effective method for simultaneous and rapid detection of HFMD virus in water environments.

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