Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, PR China.
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, 200241, PR China.
Mol Cell Probes. 2020 Oct;53:101647. doi: 10.1016/j.mcp.2020.101647. Epub 2020 Aug 8.
Feline bocavirus-1 (FBoV-1) was first discovered in Hong Kong in 2012, and studies have indicated that the virus may cause feline hemorrhagic enteritis. Currently, there is a lack of an effective and quantitative method for FBoV-1 detection. In this study, a TaqMan-based quantitative real-time PCR (qPCR) for FBoV-1 detection was established. Primers and probes were designed to target the conserved region of the FBoV-1 NS1 gene. The sensitivity analysis indicated that the minimum detection limit was 4.57 × 10 copies/μL. The specificity test revealed no cross-reaction with seven other common feline viruses, including the same species-FBoV-2 and FBoV-3. The sensitivity of this method was 100 times higher than that of conventional PCR (cPCR). The established method showed good repeatability, with the intra-assay and inter-assay coefficients of variation of 0.18%-1.00% and 0.27%-0.45%, respectively. Furthermore, the analysis of feline feces revealed that the detection rate by qPCR was 7.0% (9/128), whereas that by cPCR was 4.7% (6/128). In conclusion, the established qPCR assay can quantitatively detect FBoV-1 with a high sensitivity, high specificity, and good reproducibility, making it a promising technique for the clinical detection of and basic and epidemiological research on FBoV-1.
猫博卡病毒-1（FBoV-1）于 2012 年在香港首次发现，研究表明该病毒可能导致猫出血性肠炎。目前，缺乏有效的定量检测 FBoV-1 的方法。本研究建立了一种基于 TaqMan 的定量实时 PCR（qPCR）检测 FBoV-1 的方法。设计了针对 FBoV-1 NS1 基因保守区的引物和探针。灵敏度分析表明，最小检测限为 4.57×10 拷贝/μL。特异性试验表明，该方法与其他七种常见的猫病毒无交叉反应，包括同一种属-FBoV-2 和 FBoV-3。该方法的灵敏度比常规 PCR（cPCR）高 100 倍。该方法具有良好的重复性，内、间试验变异系数分别为 0.18%-1.00%和 0.27%-0.45%。此外，对猫粪便的分析表明，qPCR 的检测率为 7.0%（9/128），而 cPCR 的检测率为 4.7%（6/128）。总之，建立的 qPCR 检测方法可以定量检测 FBoV-1，具有高灵敏度、高特异性和良好的可重复性，是一种有前途的临床检测和 FBoV-1 基础和流行病学研究的技术。
