Li Yongdong, Yu Zhaorong, Jiao Suli, Liu Yuhui, Ni Hongxia, Wang Yong
Municipal Key Laboratory of Virology, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, PR China.
Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China.
J Virol Methods. 2020 May 26;282:113904. doi: 10.1016/j.jviromet.2020.113904.
Porcine circovirus type 3 (PCV3) is a novel member of the genus Circovirus, first detected in the United States in 2016, with subsequent reports in many countries. PCV3 infections have caused serious economic losses in the pig industry. Alternative rapid and sensitive assays for PCV3 detection are needed for clinical diagnosis, especially in laboratories not equipped with more sophisticated equipment. Here, a real-time recombinase-aided amplification assay (RAA) was developed for PCV3 detection. Specific primers and probes targeting the conserved region of the capsid gene of PCV3 were designed. The assay was performed at 39 °C for 30 min using specialized equipment. Furthermore, 36 clinical samples were used to evaluate the RAA. The analytical sensitivity of the RAA for PCV3 was 38 copies per reaction at 95% probability level, using a probit regression model. There was no cross-reactivity with other DNA viruses belonging to the Circoviridae and Parvoviridae families. The detection rate agreed with that obtained by an established real-time PCR assay with a kappa value of 1.0. Our results demonstrated that this new RAA could be used for the rapid, accurate, and sensitive detection of PCV3.
猪圆环病毒3型（PCV3）是圆环病毒属的一个新成员，2016年首次在美国被检测到，随后在许多国家都有报道。PCV3感染给养猪业造成了严重的经济损失。临床诊断需要用于PCV3检测的快速灵敏的替代检测方法，尤其是在没有配备更精密设备的实验室中。在此，开发了一种用于PCV3检测的实时重组酶辅助扩增检测法（RAA）。设计了针对PCV3衣壳基因保守区域的特异性引物和探针。该检测使用专门设备在39℃下进行30分钟。此外，使用36份临床样本对RAA进行评估。使用概率回归模型，RAA对PCV3的分析灵敏度在95%概率水平下为每个反应38个拷贝。与属于圆环病毒科和细小病毒科的其他DNA病毒无交叉反应。检测率与通过既定的实时PCR检测法获得的结果一致，kappa值为1.0。我们的结果表明，这种新的RAA可用于PCV3的快速、准确和灵敏检测。
