Development of one-step real-time reverse transcription polymerase chain reaction assays for rapid detection of porcine group C rotaviruses

作者：Chun Young Hyun; Jeong Young Ju; Park Sang Ik; Hosmillo Myra; Shin Dong Jun; Kwon Hyung Jun; Kang Shien Young; Woo Sang Kyu; Kang Mun Il; Park Su Jin^*; Cho Kyoung Oh

来源：Journal of Veterinary Diagnostic Investigation, 2010, 22(1): 74-77.

• Diagnosis
• porcine group C rotavirus
• real-time reverse transcription polymerase chain reaction
• SYBR Green
• TaqMan

Although the widespread occurrence of porcine group C rotaviruses (GCRV) is assumed, precise prevalence remains largely unknown because of the absence of reliable, specific, and rapid diagnostic methods. To detect and quantify porcine GCRV, the authors evaluated and optimized SYBR Green and TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR) assays and applied them to 108 piglet fecal samples. Using serially diluted standard RNA transcripts of porcine GCRV VP6 gene, both SYBR Green and TaqMan real-time RT-PCR assays detected as few as 1 x 10(1) genome copies/mu l (correlation coefficiency >0.99), whereas conventional RT-PCR detected 1.0 x 10(3) copies/mu l. In addition, the conventional assay detected porcine GCRV in 24% (26/108) of fecal samples, whereas the detection rates of both SYBR Green and TaqMan assays were 72% (78 of 108) and 64% (70 of 108), respectively. The current study indicated that both real-time RT-PCR assays were reliable, specific, and rapid methods for the detection of porcine GCRV in porcine fecal samples.

• 出版日期2010-1