A Simple and Sensitive RT-qPCR Technology for Rapid Detection of Porcine Reproductive and Respiratory Syndrome Virus
Hongri Zhao | Xingyu Xiao | Yajuan Sun | Yang Chen | Yongzhe Zhang | Peng Li | Hui Jin | Ying Li | Rui Yin
To establish a rapid and sensitive detection method for the porcine reproductive and respiratory syndrome virus (PRRSV), gene-specific primers and a TaqMan probe were designed based on the <i>M</i> gene of PRRSV, and a new stable fully pre-mixed reverse transcription real-time fluorescence quantitative PCR (RT-qPCR) reaction mixture was developed. A simple and rapid RT-qPCR detection method for PRRSV was developed by optimizing nucleic acid amplification conditions. The results showed that the method was able to specifically detect PRRSV without cross-reactivity with the other 11 porcine susceptible viruses. The sensitivities of the assay were 3.12 × 10<sup>0</sup> copies/μL and 10<sup>0</sup> TCID<sub>50</sub>/μL for <i>M</i> gene and virus, respectively, and the repeatability and reproducibility (relative standard deviation, CV) of the assay were less than 2.5%. Based on the new fullly pre-mixed RT-qPCR reaction mixture, the RT-qPCR detection method may provide a new, simple, and rapid method for accurately detecting PRRSV.
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