Henan Provincial Engineering Laboratory of Insect Bio-reactor and Henan Key Laboratory of Ecological Security for Water Source Region of Mid-line of South-to-North, Nanyang Normal University, 1638 Wolong Road, Nanyang, Hena, 473061, People's Republic of China.
Veterinary Laboratory, Guangzhou Zoo, Guangzhou, 510642, People's Republic of China.
BMC Vet Res. 2019 Sep 6;15(1):322. doi: 10.1186/s12917-019-2072-9.
Porcine circovirus type 3 (PCV3) is a newly emerging circovirus that might be associated with porcine dermatitis and nephropathy syndrome, reproductive failure, and cardiac and multisystemic inflammation. To aid the prevention and control of the infectious disease caused by PCV3, we developed a novel isothermal amplification assay using polymerase spiral reaction (PSR), which allows the visual detection of preserved strains and clinical samples.
This assay precisely amplified the PCV3 genome with the use of a water bath at 62 °C for 50 min. The detection limit was found to be 1.13 × 10 copies/μL by gel electrophoresis or with the use of a visible dye (an indicator comprising phenol red and cresol red). No cross-reaction with other porcine infectious viruses was observed. The detection results for 23 PCV3-positive samples by PSR were in accordance with loop-mediated isothermal amplification (LAMP) assay. The detection rate of the PSR assay for PCV3 positivity of clinical samples was 68/97, which was higher than LAMP assay (67/97).
These results indicated that the PSR assay provides an accurate and rapid method for the detection of PCV3 with high sensitivity and specificity. It is particularly suited for use in a simple laboratory setting without a thermal cycler or gel electrophoresis equipment.
猪圆环病毒 3 型（PCV3）是一种新出现的圆环病毒，可能与猪皮炎肾病综合征、生殖失败以及心脏和多系统炎症有关。为了帮助预防和控制由 PCV3 引起的传染病，我们开发了一种使用聚合酶螺旋反应（PSR）的新型等温扩增检测方法，该方法允许对保存株和临床样本进行可视化检测。
该检测方法使用 62°C 的水浴 50 分钟即可精确扩增 PCV3 基因组。通过凝胶电泳或使用可见染料（包含苯酚红和甲酚红的指示剂）检测，发现其检测限为 1.13×10 拷贝/μL。与其他猪传染性病毒无交叉反应。PSR 对 23 个 PCV3 阳性样本的检测结果与环介导等温扩增（LAMP）检测相符。PSR 检测临床样本中 PCV3 阳性的检出率为 68/97，高于 LAMP 检测（67/97）。
这些结果表明，PSR 检测方法具有高灵敏度和特异性，为 PCV3 的检测提供了一种准确、快速的方法。它特别适合在没有热循环仪或凝胶电泳设备的简单实验室环境中使用。
