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 P8, P4, P6, P9and P10 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 P8, P4, P6, P9 and P10 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.P8 was 72.5%, P4 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.