A duplex nested RT- PCR for simultaneous detection of type 2 PRRSV and PCV2 from tissue samples
Hye Kwon Kim1,†, Kwang Soo Lyoo2,†, Thi My Le Huynh3, Hyoung Joon Moon4, Van Giap Nguyen3, Bong Kyun Park5,*
1Viral Infectious Disease Research Center, Korea Research Institue of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Youseong-gu, Daejeon 305-806, Korea
2Korea Zoonosis Research Institute, Chonbuk National University, Jeonju, Jeonbuk, Korea
3Department of Veterinary Microbiology- Infectious Diseases, Faculty of Veterinary Medicine, Vietnam National University of Agriculture, Hanoi, Vietnam
4Research Unit, Green Cross Veterinary Products, Yongin, 449-903, Korea
5Department of Veterinary Medicine Virology Lab, College of Veterinary Medicine, Seoul National University, Seoul, 151-742, Korea
Correspondence to: Tel: 82-2-880-1255; Fax: 82-2-885-0263; E-mail: parkx026@snu.ac.kr
The first two authors have contributed equally to this work.
Received: March 8, 2016; Revised: June 20, 2016; Accepted: July 21, 2016; Published online: August 10, 2016.
There are high coincidences of PRRSV and PCV2 in the tissue distribution. This study established a duplex nested RT- PCR targeting the genomic RNA of type 2 PRRSV and mRNA of PCV2 in infected tissues. The method amplified discriminative bands of 347bp and 265bp specific for type 2 PRRSV and PCV2, respectively. The limit detection of the duplex nested RT- PCR was 101.5 TCID50/ml of type 2 PRRSV and 102 PCV2-infected cells/ml. The kappa statistic, measuring agreement between the methods, was 0.867, indicating a good level of agreement. This RNA-based duplex PCR approach can be another way to detect type 2 PRRSV and PCV2 simultaneously with convenience.
Keywords: type 2 PRRSV, PCV2, duplex nested RT- PCR

© 2016 The Korean Society of Veterinary Science.