TaqMan-based quantitative real-time polymerase chain reaction assay to detect porcine circovirus-like virus.

Z R Yu, Y Shao, Z Chen, Y Zhang, F Y Cheng, H Liu, Z Y Wang, J Tu, X J Song, K Z Qi
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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.

基于taqman的实时定量聚合酶链反应法检测猪圆环病毒样病毒。
本研究旨在建立一种快速、灵敏、高特异性的猪圆环病毒样病毒(PCLV) TaqMan实时定量PCR (qPCR)检测方法。根据PCLV ORF4基因的保守区设计引物和探针。该方法检测性能良好(y=-3.3257x+ 1.482, R2=0.9905),检出限为10份,灵敏度比常规PCR (cPCR)提高100倍。与其他常见病毒无交叉反应。试验内和试验间变异系数均小于1.25%。采用该方法对36份粪便标本进行了分析,阳性率为8.33%(3/36),高于cPCR检测结果。综上所述,所建立的PCLV检测方法具有较高的特异性、敏感性和重复性,可作为临床诊断和流行病学调查的工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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