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.
10.4142/jvs.2017.18.2.253
- Author:
Hye Kwon KIM
1
;
Kwang Soo LYOO
;
Thi My Le HUYNH
;
Hyoung Joon MOON
;
Van Giap NGUYEN
;
Bong Kyun PARK
Author Information
1. Infectious Disease Research Center, Korea Research Institute of Bioscience & Biotechnology, Daejeon 34141, Korea.
- Publication Type:Brief Communication
- Keywords:
duplex nested RT-PCR;
porcine circovirus type 2;
type 2 porcine reproductive and respiratory syndrome virus
- MeSH:
Circovirus;
Limit of Detection;
Methods;
Porcine Reproductive and Respiratory Syndrome*;
Porcine respiratory and reproductive syndrome virus*;
Reverse Transcriptase Polymerase Chain Reaction*;
RNA;
RNA, Messenger;
RNA-Directed DNA Polymerase*
- From:Journal of Veterinary Science
2017;18(2):253-256
- CountryRepublic of Korea
- Language:English
-
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 10(1.5) 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.
