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J. Vet. Sci. 2017; 18(2): 253-256  https://doi.org/10.4142/jvs.2017.18.2.253
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
Hye Kwon Kim1,†, Kwang Soo Lyoo2,†, Thi My Le Huynh3, Hyoung Joon Moon4, Van Giap Nguyen3, Bong Kyun Park5,*
1Infectious Disease Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 34141, Korea
2Korea Zoonosis Research Institute, Chonbuk National University, Jeonju 54596, Korea
3Department of Veterinary Microbiology-Infectious Diseases, Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Trau Quy, Gia Lam, Hanoi, Vietnam
4Research Unit, Green Cross Veterinary Products, Yongin 17066, Korea
5Department of Veterinary Medicine Virology Lab, College of Veterinary Medicine, Seoul National University, Seoul 08826, Korea
Correspondence to: Bong Kyun Park Tel: +82-2-880-1255; Fax: +82-2-885-0263; E-mail: parkx026@snu.ac.kr
Received: March 8, 2016; Revised: June 20, 2016; Accepted: July 21, 2016; Published online: June 30, 2017.
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 101.5 TCID50/mL for type 2 PRRSV and 102 infected 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.
Keywords: duplex nested RT-PCR, porcine circovirus type 2, type 2 porcine reproductive and respiratory syndrome virus


© 2017 The Korean Society of Veterinary Science.

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