Wang Jianchang, Zhang Yongning, Wang Jinfeng, Liu Libing, Pang Xiaoyu, Yuan Wanzhe
Center of Inspection and Quarantine, Hebei Entry-Exit Inspection and Quarantine Bureau, Shijiazhuang, Hebei 050051, China.
Institute of Animal Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100176, China.
J Virol Methods. 2017 Oct;248:177-180. doi: 10.1016/j.jviromet.2017.07.007. Epub 2017 Jul 23.
Porcine circovirus 3 (PCV3) is a novel circovirus that was associated with porcine dermatitis and nephropathy syndrome, reproductive failure, and multisystemic inflammation. The objective of this study was to develop a rapid, simple, specific and sensitive TaqMan-based real-time PCR assay for PCV3 detection. Specific primers and probe were designed for the cap gene of PCV3 within the conserved region of viral genome. The assay was highly specific for PCV3, without cross-reactions with other non-targeted pig viruses. The detection limit of this assay was 10 copies. The assay had an efficiency of 95.7%, a regression squared value (R) of 0.994 and showed a linear range of 10-10 copies PCV3 DNA per reaction. The assay was also very reproducible, with the intra- and inter-assay coefficient of variation less than 2.0%. For the 112 archived clinical samples collected from 2014 to March 2017, the PCV3 positive ratio was 12.5% (14/112) with the real-time PCR. The presence of the PCV3 dated back to at least 2014 in China and samples collected in 2017 had the highest PCV3 positive ratio (46.7%, 7/15). The real-time PCR assay could be used for detection of PCV3 in epidemiological and pathogenesis studies.
猪圆环病毒3型（PCV3）是一种新型圆环病毒，与猪皮炎肾病综合征、繁殖障碍及多系统炎症相关。本研究的目的是建立一种快速、简便、特异且灵敏的基于TaqMan的实时荧光定量PCR检测方法用于PCV3检测。针对PCV3病毒基因组保守区域的cap基因设计了特异性引物和探针。该检测方法对PCV3具有高度特异性，与其他非目标猪病毒无交叉反应。该检测方法的检测限为10个拷贝。该检测方法的效率为95.7%，回归平方值（R）为0.994，每个反应中PCV3 DNA的线性范围为10 - 10拷贝。该检测方法的重复性也非常好，批内和批间变异系数均小于2.0%。对于2014年至2017年3月收集的112份存档临床样本，实时荧光定量PCR检测显示PCV3阳性率为12.5%（14/112）。PCV3在中国至少可追溯到2014年，2017年收集的样本中PCV3阳性率最高（46.7%，7/15）。该实时荧光定量PCR检测方法可用于PCV3的流行病学和发病机制研究。
