猪捷申病毒与猪圆环病毒2型双重PCR检测方法的建立及初步应用

Establishment and Application of Duplex PCR Assay for the Detection of Porcine Teschovirus and Porcine Circovirus Type 2

为建立可同时检测猪捷申病毒(PTV)与猪圆环病毒2型(PCV2)的双重PCR方法,本研究根据GenBank登录的相关病毒基因序列,选择保守区域设计引物,经过反应条件的优化,建立了可同时检测以上2种病毒的PCR诊断方法,扩增片段长度分别为187 bp(PTV)、120 bp(PCV2)。通过实验证明该方法具有良好的特异性和较高的敏感性,对PTV、PCV2核酸检测最低浓度分别为2.8×10-2 ng和6.6×10-3 ng。应用该方法对43份临床样品进行检测发现,PTV阳性率为23%,PCV2阳性率为38%,PTV与PCV2共感染率为16%。该方法的成功建立,为快速高效地检测以上2种病毒提供有力手段。

This experiment was conducted to establish a duplex PCR method whichwas developed for detecting the porcine teschovirus （ PTV ） and porcine circovirus type 2 （ PCV2 ） with a set of specific primers designed according to the published viral genome sequences. The PCR products for each virus were 187 bp （PTV） and 120 bp （PCV2） , with the detection limit of 2.8 ×10^-2 ng and 6.6 × 10^-3 ng, respectively. The method was used to detect 43 clinical samples. Among the samples, PTV percentage was 23%, PCV2 positive percentage was 38% , and co -infection positive percentage was 16%. This duplex PCR assay having been successfully established was proved to be a sensitive, specific method for rapid detection of the two viruses.