TaqMan-based real-time polymerase chain reaction assay for specific detection of bocavirus-1 in domestic cats. (October 2020)

• Record Type: Journal Article
• Authors: Wang, Yong; Sun, Jianfei; Guo, Xu; Zhang, Da; Cui, Yongqiu; Li, Wei; Liu, Guangqing; Li, Yongdong; Jiang, Shudong
• Is Part Of: Molecular and cellular probes. Volume 53(2020)
• Publication Date: 2020-10
• Subjects: Feline bocavirus-1 -- NS1 gene -- TaqMan -- Real-time polymerase chain reaction; Molecular probes -- Diagnostic use -- Periodicals; Pathology, Cellular -- Technique -- Periodicals; Cell Biology -- Periodicals; Molecular Biology -- Periodicals; Sondes moléculaires -- Utilisation diagnostique -- Périodiques; Cytopathologie -- Technique -- Périodiques; 572
• DOI: 10.1016/j.mcp.2020.101647
• Languages: English
• ISSNs: 0890-8508

Abstract

Abstract: 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 1 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. Highlights: The study establishes a TaqMan qPCR method for Feline Bocavirus-1 detection. The assay has high sensitivity, specificity and reliability. The study provides a tool that could be beneficial for clinical diagnostics of Feline Bocavirus-1 infection as well as future research on the virus.