Development of one-step real-time reverse transcription polymerase chain reaction assays for rapid detection of porcine group C rotaviruses.
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/microl (correlation coefficiency >0.99), whereas conventional RT-PCR detected 1.0 x 10(3) copies/microl. 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.
Chun
Young-Hyun
YH
Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology, Jeongeup 580-185, South Korea.
Jeong
Young-Ju
YJ
Park
Sang-Ik
SI
Hosmillo
Myra
M
Shin
Dong-Jun
DJ
Kwon
Hyung-Jun
HJ
Kang
Shien-Young
SY
Woo
Sang-Kyu
SK
Kang
Mun-Il
MI
Cho
Kyoung-Oh
KO
D000818:Animals; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012401:Rotavirus / Q000145:classification / Q000302:isolation & purification*; D012400:Rotavirus Infections / Q000662:veterinary* / Q000821:virology; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology; D013997:Time Factors