Application of GeXP multiple gene expression heredity analysis system in detection of nine encephalitis related viruses

Abstract

The invention belongs to the biotechnological application filed, and relates to simultaneous detection and typing of the infection of nine encephalitis viruses (comprising banna viruses, GI encephalitis B viruses, GIII encephalitis B viruses, tick-borne encephalitis viruses, Tahyna viruses, Liaoning viruses, Kyasanur forest fever viruses, Sindbis viruses and Yunnan orbiviruses) of viral encephalitis patient specimens in disease prevention control mechanisms at all levels, sentinel point hospitals and the like. The nucleotide sequences of nine encephalitis related virus strains are downloaded from NCBI, the pathogen relative conservation region is determined through literature consulting and multiple sequence alignment, and multiple specific primers are designed. The single tube 13-plex multiplex PCR test is carried out to detect nine encephalitis virus conservation regions, and the consumption time of the whole reaction is less than 2h. According to the invention, the non-typing disadvantage of routine single-tube multiple fluorescent qualitative PCR detection is overcome, the disadvantages comprising complex operation, long time, high cost and the like of routine chip detection methods are overcome, a new thought is provided for an encephalitis virus typing technology, the characteristics comprising high specificity, high sensitivity and rapidness of the GeXP multiple gene expression heredity analysis system provide a strong technological support for the rapid and accurate screening and typing of the encephalitis viruses, and the GeXP multiple gene expression heredity analysis system is of important significance to the encephalitis syndrome patient infection pathogen spectrum and molecular epidemiology investigation in China.

Description

field of invention [0001] The invention belongs to the field of biotechnology application, and relates to nine kinds of encephalitis-associated viruses (including Banna virus, GI Japanese encephalitis virus, GIII B virus, etc.) Simultaneous detection and typing of encephalovirus, tick-borne encephalitis virus, Tahyna virus, Liaoning virus, Kosanur forest fever virus, Sindbis virus, and Yunnan orbivirus) infection. Specifically, the nucleotide sequences of nine representative strains of encephalitis viruses were downloaded from NCBI, and the relatively conserved regions of the pathogen were determined by consulting the literature and multiple sequence comparisons, and multiple specific primers were designed. A single-tube multiplex (13-fold) PCR was performed to detect the conserved regions of 9 encephalitis viruses, and the entire reaction took less than 2 hours. This patent overcomes the shortcomings of virus culture method, direct fluorescent antibody method and chip detect...

AI Technical Summary

Problems solved by technology

In recent years, various molecular biology diagnostic methods based on PCR technology have been widely used and developed, including SYBR Green I real-time PCR and TaqMan PCR, etc., but they are basically used for single virus detection (such as Japanese encephalitis). inflammatory virus), it is difficult to make an accurate diagnosis of unknown pathogenic infection cases

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