猪生殖与呼吸综合征病毒RT-LAMP检测方法的建立

Development of a RT-LAMP assay for detection of porcine reproductive and respiratory syndrome virus

针对美洲型猪生殖与呼吸综合征病毒(PRRSV)保守的N蛋白基因设计了一套RT-LAMP的检测引物,通过优化反应条件,建立了一种检测PRRSV的RT-LAMP诊断方法。特异性和敏感性试验结果显示,该方法能够特异地检测美洲型PRRSV,其最低检测限为10个拷贝,而RT-PCR的最低检出限为1×104个拷贝。分别用建立的RT-LAMP法与RT-PCR法对临床50份样品进行检测,结果二者的符合率在96%以上,呈现很好的一致性。表明,建立的RT-LAMP法可应用于PRRSV的快速检测。

A set of reverse transcription loop-mediated isothermal amplification（RT-LAMP） detection primers were designed according to the conserved N protein gene of porcine reproductive and respiratory syndrome virus（PRRSV） American type. Through optimization of reaction condition,a RT-LAMP assay for clinical diagnosis of PRRSV was established under isothermal conditions. The assay could detect specifially PRRSV American type and achieved theoretically a sensitivity of 10 plasmid molecules,whereas that of RT-PCR was 1×10^4copies. The coincidence rate of RT-LAMP with RT-PCR was over 96% based on detection of 50 clinical samples,indicating that there was high correlation between RT PCR and RT-LAMP. The results showed that the RT-LAMP assay possessed potential for rapid detection of PRRSV.