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J Virol Methods. 2017 Dec;250:11-16. doi: 10.1016/j.jviromet.2017.09.021. Epub 2017 Sep 21.

Multiplex real-time polymerase chain reaction for the differential detection of porcine circovirus 2 and 3.

Author information

1
College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea.
2
Animal Disease Diagnostic Division, Animal and Plant Quarantine Agency, Gimcheon 39660, Republic of Korea.
3
College of Veterinary Medicine Konkuk University, Seoul 05029, Republic of Korea.
4
College of Veterinary Medicine & Animal Disease Intervention Center, Kyungpook National University, Daegu 41566, Republic of Korea. Electronic address: parkck@knu.ac.kr.

Abstract

A multiplex quantitative real-time polymerase chain reaction (mqPCR) assay was developed for the rapid and differential detection of porcine circovirus 2 (PCV2) and PCV3. Each of the capsid genes of PCV2 and PCV3 were amplified using specific primers and probe sets, while no other porcine pathogen genes were detected. Limit of detection of the assay was below 50 copies of the target genes of PCV2 and PCV3, and was comparable to that of previously described methods The assay showed high repeatability and reproducibility, with coefficients of intra-assay and inter-assay variation of less than 4.0%. Clinical evaluation using tissue samples from a domestic pig farm showed that PCV2 and PCV3 co-circulated at the farm. Moreover, singular infection rates of PCV2 or PCV3 were 21.7% (10/46) or 6.5% (3/46), respectively, while the co-infection rate of PCV3 with PCV2 was 28.3% (13/46). PCV3 DNA was detected by the mqPCR in respiratory diseased piglet tissue samples and aborted fetal tissue samples, suggesting that PCV3 infection is associated with porcine respiratory disease and reproductive failure in the pig farm. This mqPCR method is a rapid and reliable differential diagnostic tool for the monitoring and surveillance of PCV2 and PCV3 in the field.

KEYWORDS:

Capsid gene; Multiplex; PCV3; Porcine circovirus 2; Real-time PCR

PMID:
28941615
DOI:
10.1016/j.jviromet.2017.09.021
[Indexed for MEDLINE]

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