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. 2014 Jun:201:13-9.
doi: 10.1016/j.jviromet.2014.01.016. Epub 2014 Feb 18.

The application of a duplex reverse transcription real-time PCR for the surveillance of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2

Chia-Yi Chang  1 , Ming-Chung Deng  1 , Fun-In Wang  2 , Hsiang-Jung Tsai  3 , Chia-Huei Yang  1 , Chieh Chang  1 , Yu-Liang Huang  4
Affiliations

The application of a duplex reverse transcription real-time PCR for the surveillance of porcine reproductive and respiratory syndrome virus and porcine circovirus type 2

Chia-Yi Chang et al. J Virol Methods. 2014 Jun.

Abstract

The porcine respiratory disease complex (PRDC) is the most common disease in commercial pork production worldwide. Porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), the most important agents of PRDC, usually co-infect in the same pigs. In order to survey the prevalence of PCV2 and PRRSV in pigs of various ages, a duplex reverse transcription real-time PCR (DRT-rPCR) was developed and applied in the present study. The DRT-rPCR did not cross-react with 10 swine viruses other than PCV2 and PRRSV, with detection limits of 1 TCID50/ml for PCV2 and 6.3 TCID50/ml for PRRSV. Surveillance using DRT-rPCR together with serology revealed that in the five farms studied, pigs were most susceptible to PRRSV at 6-14 weeks of age, whereas susceptibility to PCV2 varied by the management system but was mostly at 10-14 weeks of age. Cross analysis of viral loads versus antibody titers revealed that PCV2 load was affected negatively by anti-PCV2 ORF2 antibody, which constituted the most important non-infectious factor affecting the development of PMWS. These results indicated that DRT-rPCR was developed and applied successfully to the surveillance of PCV2 and PRRSV in the field.

Keywords: Duplicate reverse transcription real-time PCR; Porcine circovirus type 2; Porcine reproductive and respiratory syndrome virus; Surveillance.

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