The Sequence The VP6 Gene And Preliminary Application The TaqMan Based Real Time Quantitative PCR For Porcine Rotavirus

Porcine rotavirus belongs to the Reoviridae family,Rotavirus genus.The virus is one of the major pathogens which can cause life-threatening diarrhea to the piglets.The piglets infected rotavirus can cause death ration nearly 20 percentage in one to four week-age pigs.Rotavirus diarrhea's typically clinical manifestation is serious diarrhea,that make the piglets dead easily with secondary infection with other infectious diseases.In this study,the VP6 gene of the porcine rotavirus isolates RV-FJ2014-01 strains was cloned by RT-PCR methods with specific primers.The results demonstrated that the full-length of the VP6 gene had 1 194 nucleotides,encoding 397 amino acids.The RV-FJ2014-01 strains nucleotides share highest to the pig/China/NMTL/2008/G9P(GenBank accession number JF781162)is 97.7%with homology comparison.Also shared with human-origin rotavirus TGE13-77(GenBank accession number LC000537)at 90.3%with homology comparison,and 90.5%homology comparison with the giant panda strain(GenBank accession number GU188283).In order to develop a more sensitive and specific diagnostic test for rotavirus by using the real time quantitative RT-PCR technology,TaqMan based real time quantitative RT-PCR technology had reported preciously for many pathogens in animal virus,with more rapidity,sensitivity,reproducibility and less risk of contamination.According by the characterization of the VP6 gene of the porcine rotavirus RV-FJ2014-01 strains.We established the TaqMan based real time quantitative RT-PCR method for detecting the rotavirus infection status,with the correlation value for the amplification at 0.999.Statistics of TaqMan based real time PCR within the groups and between the groups is 0.25%?1.26%and 0.31%?1.49%.In conclusion,the results in this study can lay good foundation for the epidemiological investigation;enrich the genomic information and rapid diagnosis method of porcine rotavirus.