title: Development of one-step real-time reverse transcription polymerase chain reaction assays for rapid detection of porcine group C rotaviruses.
abstract: 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.
authors.affiliation: Bioindustry Research Center, Korea Research Institute of Bioscience and Biotechnology, Jeongeup 580-185, South Korea.
authors.forename: Young-Hyun
authors.initials: YH
authors.lastname: Chun
authors.forename: Young-Ju
authors.initials: YJ
authors.lastname: Jeong
authors.forename: Sang-Ik
authors.initials: SI
authors.lastname: Park
authors.forename: Myra
authors.initials: M
authors.lastname: Hosmillo
authors.forename: Dong-Jun
authors.initials: DJ
authors.lastname: Shin
authors.forename: Hyung-Jun
authors.initials: HJ
authors.lastname: Kwon
authors.forename: Shien-Young
authors.initials: SY
authors.lastname: Kang
authors.forename: Sang-Kyu
authors.initials: SK
authors.lastname: Woo
authors.forename: Mun-Il
authors.initials: MI
authors.lastname: Kang
authors.forename: Kyoung-Oh
authors.initials: KO
authors.lastname: Cho
mesh_terms: 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