GeXP system combined with multiplex PCR technology for detection of diarrhea viruses

Abstract

Objective GenomeLab Genetic Analysis System (GeXP) and multiplex-PCR were combined to detect Noroviruse I、II, Astrovirus and Rotavirus. Methods The sequences of these viruses were obtained from Genbank of NCBI, and specific primer pairs were designed by GeXP eXpress Profiler. After optimization, the GeXP system could amplify specific fragments of each virus. The sensitivity and specificity of multiple PCR assay was also evaluated.Optimized assay was further validated with 120 clinical specimens collected from the CDC. Results These primer pairs were successfully used for detecting these viruses at the level of 5×103copies/ μl without cross reaction. The sensitivity, specificity and efficiency of GeXP assay was 100%, 99.53% and 99.62%, respectively. Comparing to Real-time PCR, the consistence rate reached 99.51% in 120 clinical samples. Conclusions The combined detection of GeXP and multiplex PCR assay could be a high-throughput, rapid and highly specific and sensitive method used in the screening of the diarrhea viruses. Key words: Diarrhea viruses; GeXP; Multiplex-PCR