Sun Pei, Ye Yumeng, Li Yeqiu, Cui Yongqiu, Zhou Tianhong, Li Yongdong, Wang Yong
Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036 People's Republic of China.
New Ruipeng Hefei Tianhong Pet Hospital, Hefei, 230036 People's Republic of China.
3 Biotech. 2021 Nov;11(11):472. doi: 10.1007/s13205-021-03031-z. Epub 2021 Oct 23.
In the study, we established a hydrolysis probe-based real-time polymerase chain reaction (PCR) assay to rapidly detect Canine circovirus (CanineCV) DNA in faecal samples. We designed a pair of specific primers and one probe targeting in CanineCV, and sensitivity, specificity, and repeatability tests were performed to evaluate the efficacy of the assay. The assay showed high sensitivity and a minimum detection limit of 8.42 × 10 copies/μL, which is 1000-fold more sensitive compared to traditional PCR. The method was also highly specific, without cross-reaction with other common canine viruses. Moreover, the assay showed high repeatability, and the mean intra-assay and inter-assay coefficients of variation were 0.26 and 0.36%, respectively. The results of the detection of clinical samples showed that the positive detection rate of CanineCV was 14.04% (8/57). Notably, 8% of clinical samples were co-infected with other canine pathogens. In conclusion, the establishment of a hydrolysis probe-based real-time PCR method provides a fast, sensitive, specific, reliable, and repeatable method for CanineCV detection.
The online version contains supplementary material available at 10.1007/s13205-021-03031-z.
在本研究中，我们建立了一种基于水解探针的实时聚合酶链反应（PCR）检测方法，用于快速检测粪便样本中的犬圆环病毒（CanineCV）DNA。我们设计了一对针对CanineCV的特异性引物和一个探针，并进行了灵敏度、特异性和重复性测试，以评估该检测方法的有效性。该检测方法显示出高灵敏度，最低检测限为8.42×10拷贝/μL，比传统PCR灵敏1000倍。该方法也具有高度特异性，与其他常见犬病毒无交叉反应。此外，该检测方法显示出高重复性，批内和批间变异系数的平均值分别为0.26%和0.36%。临床样本检测结果显示，CanineCV的阳性检出率为14.04%（8/57）。值得注意的是，8%的临床样本与其他犬病原体共同感染。总之，基于水解探针的实时PCR方法的建立为CanineCV检测提供了一种快速、灵敏、特异、可靠且可重复的方法。
在线版本包含可在10.1007/s13205-021-03031-z获取的补充材料。
