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Published on: November 1, 2018
TaqMan-based quantitative real-time polymerase chain reaction assay to detect porcine circovirus-like virus
1Anhui Province Engineering Laboratory for Animal Food Quality and Bio-safety, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China.
Abstract:
The aim of this study was to develop a rapid, sensitive and highly specific TaqMan quantitative real-time polymerase chain reaction PCR (qPCR) assay for porcine circovirus-like virus (PCLV). The primers and probe were designed based on the conserved regions of the PCLV ORF4 gene. The assay has a good detection performance (y=-3.3257x+ 1.482, R2=0.9905), with a limit of detection of 10 copies, which was 100 times more sensitive than conventional PCR (cPCR). No cross-reactivity was observed with other common viruses. The intra- and inter-assay coefficients of variation were less than 1.25%. 36 fecal samples were analyzed using this method, detecting a positivity rate of 8.33% (3/36) that was higher than the cPCR detected. In summary, the established assay for PCLV detection has high specificity, sensitivity, and reproducibility and can be used as a tool for clinical diagnosis and epidemiological investigation.
