Establishment and application of a one-step real-time fluorescence quantitative PCR assay for bovine respiratory syncytial disease
To establish a rapid and simple method for the detection of bovine respiratory syncytial virus(BRSV),we designed specific primers and probes based on the sequence of the conserved region of the BRSV M gene,and established a one-step real-time fluorescence quantitative PCR assay for BRSV detection by optimizing the reaction conditions,and validated the sensitivity,specificity and stability of the assay.Finally,the method was also used to test clinical samples.The results showed that this study successfully established a BRSV fluores-cence quantitative assay with good specificity and specific amplification only for BRSV,with high sensitivity at a minimum of 10 copies/μL,with good stability with low intra-group and inter-group coefficients of variation.The detection rate of 94 clinical samples collected in the Ningxia region using the established BRSV one-step real-time fluorescence quantitative PCR assay,the positive rate was 5.3%(5/94).The above results indicated that the method established in this study might serve as a powerful technical tool for rapid diagnosis of BRSV.
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