Development new multiplex reverse transcription quantitative real time polymerase reaction to detect human norovirus genogroups I and II

Norovirus (NV) is considering a highly contagious virus due to the increase in the number of cases of the virus in recent years. Therefore, there is a need to develop rapid, specific and sensitive molecular detection and diagnosis methods. Using the data from the National Center of Bioinformatics Information (NCBI), the sequence of more than 1000 Norovirus genogroupII genome and 39 genome of geno group I were chosen to select a common conserve region for all genotypes within the first group and the second group suitable for the design of primers and probes. The design of the first area of the junction of Open Reading Frame one (ORF) and the open reading frame two (ORF2), and the primers and probes were tested on samples of compared to the primers and using one step reverse transcription real time PCR. The kit designed for multiplex reverse reaction possess no overlap or interaction in the components of primers and probes of the two genogroups, also it is superior on the monoplex mixture of the previous kit, Norovirus being diagnosed in 45% of samples by primers and probes designed in this study, 10% of the samples were belong to the genogroupI and 35% of the samples belong to the genogroupII in comparing with virus results that tested with monoplex previous primers and probes accounted 44% , the genogroupI represented 10% and genogroupII II represented 34% of the tested samples. The use of the multiplex kit contributes to shortening the time and the largest number of samples and reduce amount of the solutions used, more than the monoplex reaction, as it should be performed separately for each genetic group.