Establishment and preliminary application of duplex fluorescence quantitative PCR for porcine circoviruses type 2 and type 3
Yong-Yu Gao,
Qian Wang,
Shuang Zhang,
Jian Zhao,
Di Bao,
Han Zhao,
Kai Wang,
Gui-Xue Hu,
Feng-Shan Gao
Affiliations
Yong-Yu Gao
College of Animal Medicine, Jilin Agricultural University, Changchun, 130118, China
Qian Wang
The Third Affiliated Hospital of Changchun University of Traditional Chinese Medicine, Changchun, Jilin, 130117, China
Shuang Zhang
College of Animal Medicine, Jilin Agricultural University, Changchun, 130118, China
Jian Zhao
ChangChun Sino Biotechnology CO., LTD., Changchun, Jilin, 130012, China
Di Bao
College of Animal Medicine, Jilin Agricultural University, Changchun, 130118, China
Han Zhao
College of Animal Medicine, Jilin Agricultural University, Changchun, 130118, China
Kai Wang
College of Animal Medicine, Jilin Agricultural University, Changchun, 130118, China; Corresponding author. Department of Preventive Veterinary Medicine, College of Animal Medicine, Jilin Agricultural University, Xincheng Street 2888, Changchun, Jilin 130118, China.
Gui-Xue Hu
College of Animal Medicine, Jilin Agricultural University, Changchun, 130118, China; Corresponding author. Department of Preventive Veterinary Medicine, College of Animal Medicine, Jilin Agricultural University, Xincheng Street 2888, Changchun, Jilin, 130118, China.
Feng-Shan Gao
College of Life and Health, Dalian University, Dalian, 116622, China; The Dalian Animal Virus Antigen Epitope Screening and Protein Engineering Drug Developing Key Laboratory, Dalian, 116622, China; Corresponding author. Xuefu Street 10, Department of Bioengineering, College of Life and Health, Dalian University, Dalian, Liaoning, 116622, China.
Porcine circovirus types 2 (PCV2) and 3 (PCV3) are the two most prevalent porcine circoviruses in China, all of which can infect swine herds and cause serious diseases. To detect coinfection with PCV2 and PCV3, primers and probes for duplex PCV2 and PCV3 real-time PCR were designed to target their cap genes based on the constructed plasmids pUC57-PCV2 and pUC57-PCV3. The established duplex PCV2 and PCV3 real-time PCRs were specific to PCV2 and PCV3 and showed no cross-reactions with other porcine viral pathogens. The limit of detection was 5 and 50 copies for the PCV2 and PCV3 plasmids, respectively. The intra- and interassay repeatability had coefficients of variation below 3 %. The established methods were used to analyze clinical samples from Liaoning and Jilin provinces of China. The coinfection rates of PCV2 and PCV3 in pigs extensively fed in Liaoning and Jilin, large-scale farmed pigs in Liaoning and large-scale farmed pigs in Jilin were 15.0 % (6/40), 36.7 % (11/30) and 35.4 % (62/175), respectively. This study established a useful duplex PCV2 and PCV3 real-time PCR method that can be used for the detection of PCV2 and PCV3 in local clinical samples.
