Bian Zhibiao, Cai Rujian, Jiang Zhiyong, Song Shuai, Li Yan, Chu Pinpin, Zhang Kunli, Yang Dongxia, Gou Hongchao, Li Chunling
Institute of Animal Health, Guangdong Academy of Agricultural Sciences, Guangzhou, China.
Guangdong Provincial Key Laboratory of Livestock Disease Prevention, Guangzhou, China.
Front Vet Sci. 2021 Sep 3;8:726723. doi: 10.3389/fvets.2021.726723. eCollection 2021.
Since 2016, a novel porcine circovirus, PCV3, has been infecting pigs, causing significant economic losses to the pig industry. In recent years, the infection rate of PCV3 has been increasing, and thus rapid and accurate detection methods for PCV3 are essential. In this study, we established a novel probe-based single multiple cross displacement amplification (P-S-MCDA) method for PCV3. The method was termed as P-S-MCDA. The P-S-MCDA uses seven primers to amplify the capsid gene, and the assay can be performed at 60°C for 30 min, greatly shortening the reaction time. The results of P-S-MCDA can not only be monitored in real time through fluorescence signals but also be determined by observing the fluorescence of the reaction tubes using a smartphone-based cassette. This method demonstrated good specificity and the same sensitivity as qPCR, with a minimum detection limit of 10 copies. In 139 clinical samples, the coincidence rate with qPCR was 100%. The P-S-MCDA can be widely applied in PCV3 detection in laboratories or in rural areas.
自2016年以来，一种新型猪圆环病毒PCV3一直在感染猪，给养猪业造成了重大经济损失。近年来，PCV3的感染率一直在上升，因此快速准确的PCV3检测方法至关重要。在本研究中，我们建立了一种新型的基于探针的单多重交叉置换扩增（P-S-MCDA）方法用于检测PCV3。该方法被称为P-S-MCDA。P-S-MCDA使用七条引物扩增衣壳基因，该检测可在60°C下进行30分钟，大大缩短了反应时间。P-S-MCDA的结果不仅可以通过荧光信号实时监测，还可以使用基于智能手机的暗盒观察反应管的荧光来确定。该方法显示出良好的特异性，与qPCR具有相同的灵敏度，最低检测限为10个拷贝。在139份临床样本中，与qPCR的符合率为100%。P-S-MCDA可广泛应用于实验室或农村地区的PCV3检测。
