title: Duplex nested reverse transcriptase polymerase chain reaction for simultaneous detection of type 2 porcine reproductive and respiratory syndrome virus and porcine circovirus type 2 from tissue samples.
abstract: There are high levels of co-incidence of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) in porcine tissue. This study established a duplex nested reverse transcriptase polymerase chain reaction (RT-PCR) method that targets the genomic RNA of type 2 PRRSV and the mRNA of PCV2 in infected tissues. The method amplified discriminative bands of 347 bp and 265 bp specific for type 2 PRRSV and PCV2, respectively. The limits of detection of the duplex nested RT-PCR were 101.5TCID50/mL for type 2 PRRSV and 102infected cells/mL for PCV2. The kappa statistic, which measures agreement between methods, was 0.867, indicating a good level of agreement. This RNA-based duplex RT-PCR approach can be another way to detect type 2 PRRSV and PCV2 simultaneously and with improved convenience.
authors: Infectious Disease Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 34141, Korea.
authors: Hye Kwon
authors: HK
authors: Kim
authors: Korea Zoonosis Research Institute, Chonbuk National University, Jeonju 54596, Korea.
authors: Kwang Soo
authors: KS
authors: Lyoo
authors: Department of Veterinary Microbiology-Infectious Diseases, Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Trau Quy, Gia Lam, Hanoi, Vietnam.
authors: Thi My Le
authors: TML
authors: Huynh
authors: Research Unit, Green Cross Veterinary Products, Yongin 17066, Korea.
authors: Hyoung Joon
authors: HJ
authors: Moon
authors: Department of Veterinary Microbiology-Infectious Diseases, Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Trau Quy, Gia Lam, Hanoi, Vietnam.
authors: Van Giap
authors: VG
authors: Nguyen
authors: Department of Veterinary Medicine Virology Lab, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea.
authors: Bong Kyun
authors: BK
authors: Park
mesh_terms: D000818:Animals; D018173:Circoviridae Infections / Q000175:diagnosis* / Q000821:virology; D017925:Circovirus / Q000235:genetics*; D060085:Coinfection / Q000175:diagnosis / Q000662:veterinary* / Q000821:virology; D057230:Limit of Detection; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D019318:Porcine Reproductive and Respiratory Syndrome / Q000175:diagnosis* / Q000821:virology; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis* / Q000821:virology
keywords: duplex nested RT-PCR; porcine circovirus type 2; type 2 porcine reproductive and respiratory syndrome virus