关键词: 多重RT-PCR, PEDV, TGEV, GAR

Abstract: To develop a multiplex reverse transcription polymerase chain reaction (mutiplex RT-PCR) for detection of porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV) and porcine group A rotavirus (GAR), three pairs of primers targeting the M gene, N gene ,VP7 gene of PEDV, TGEV, GAR were designed respectively. Using the three pairs of primes, a multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) was developed and optimized. And we compared the results of multiplex RT-PCR with routine RT-PCR in the field trail. In the laboratory test, we found that the detection limit of multiplex RTPCR is 35 pg RNA of TGEV-PEDV-GAR vaccine. And in the field trail, 75 fecal specimens collected from pigs with diarrhea in the central area of China were simultaneous tested by the multiplex RT-PCR and routine RT-PCR. The relative sensitivity and specificity of multiplex RTPCR were evaluated. The results suggested that this assay was equal in quality to routine RT-PCR assays (sensitivitys were 92%, 100%, 100% for PEDV, TGEV, GAR respectively; specificity was 100% for all 3 kind of viruses). The results indicated that the multiplex RT-PCR with high sensitivity and specificity provided a new and alternative tool for the detection of PEDV, TGEV and GAR.

Key words: multiplex RT-PCR, PEDV, TGEV, GAR