数字农科院2.0

Taqman-Based Real-Time Polymerase Chain Reaction Assay For Specific Detection Of Bocavirus-1 In Domestic Cats

文献类型： 外文期刊

作者： Wang, Y; Sun, JF; Guo, X; Zhang, D; Cui, YQ; Li, W; Liu, GQ; Li, YD; Jiang, SD

作者机构：

关键词： Feline Bocavirus-1; Ns1 Gene; Taqman; Real-Time Polymerase Chain Reaction

期刊名称： MOLECULAR AND CELLULAR PROBES

ISSN： 0890-8508

年卷期： 2020 年 53 卷

页码：

收录情况： JCR（2021版）

摘要： 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 NSI gene. The sensitivity analysis indicated that the minimum detection limit was 4.57 x 10(1) copies/mu L. 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.

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