Simultaneous detection of genotype P of human group a rotaviruses by multiplex RT-PCR

Objective To establish a convenient method for simultaneously determining P-genotype of human group A rotavirus using a multiplex polymerase chain reaction.Methods 6 oligonucleotide primers which were type-specific respectively for P[8],P[4],P[6],P[9] and P[10]P-genotype of human group A rotavirus were used.The amplified product of 346、bp,484、bp,268、bp,392、bp and 584、bp represented genotype P[8],P[4],P[6],P[9] and P[10] respectively.The concentration of primers for multiplex PCR was optimized.Results All five P-genotypes of human group A rotavirus could be typed simultaneously from a mixture of 5 standard strains and the sensitivity of the assay is 0.1、ng/μl.40 clinical samples were tested.P[8] was 72.5%,P[4] was 12.5%.Conclusion The multiplex PCR appears to be rapid,sensitive and specific method for detecting human group a rotavirus' genotype P and may be applied to clinical diagnosis and laboratory work.