Method for quickly detecting main genetype of enterovirus in water environment

Abstract

The invention discloses a method for quickly detecting a main genetype of enterovirus in water environment. The method is characterized by utilizing a general primer of the enterovirus to carry out semi-nested PCR (Polymerase Chain Reaction) amplification, and combining with a DGGE analyzing method to identify a gene type of the main enterovirus in the water environment according to conservative property of nucleic acid of the enterovirus RNA at a section of 5' noncoding region. The semi-nested PCR is adopted, so that the sensitivity of an enterovirus detection technology in the water environment is greatly improved, and favorable specificity is realized; and the DGGE method is combined to separate PCR amplification sections of different genetype enteroviruses, so that the time for analyzing and identifying the main genetype of the enterovirus in the water environment is shortened, expensive cost generated by large flux sequence measurement is avoided, and the enterovirus has no cross reaction with other common pathogenic microorganisms in the environment. The method is applicable to detection of various water samples such as surface water and waste water.

Description

technical field [0001] The invention relates to a rapid detection method for the main genotypes of enteroviruses in water environment. The method is based on the general primers of enterovirus RNA conservative sequences, by adding GC splints to the primers, and using semi-nested PCR-DGGE technology to detect enteroviruses The main genotype of the virus is suitable for the rapid detection of the main genotype of the enterovirus in the water 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

[0004] Existing methods for genotyping enteroviruses in water environments are rare, and high-throughput pyrosequencing is often used in the analysis of major genotypes, which is very expensive

Although PCR-DGGE technology has been applied to the detection of bacterial community diversity and kinship identification of environmental microorganisms, there is currently no effective method for genotyping enteroviruses in water environments.

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