反转录环介导等温扩增技术快速检测猪水疱病毒方法的建立

Method establishment of a reverse transcription loop-mediated isothermal amplification to detect Swine vesicular disease virus

为了建立一种针对猪水泡病毒(SVDV)的反转录环介导等温扩增技术(RT-LAMP),用于减少猪水泡病与口蹄疫因临床症状相似而造成的误诊,试验针对SVDV VP1基因序列设计4个LAMP引物,其中外侧引物用于SVDV PCR检测,并利用RT-LAMP和RT-PCR方法进行敏感性试验、特异性试验以及临床样品检测。结果表明:针对SVDV的RT-LAMP检测方法的敏感性是RT-PCR的10倍;RT-LAMP方法的特异性良好,与口蹄疫病毒无交叉反应;两种方法对临床样品检测的符合率为100%。说明试验建立的RT-LAMP方法具有敏感性高、特异性强、快速等特点,适合实验室和临床对SVDV的快速诊断。

The aim of the present study was to develop a method of reverse transcriptase loop-mediated isothermal amplification(RT-LAMP)assay for rapidly detecting Swine vesicular disease virus(SVDV)to distinguish between Foot-and-mouth disease virus(FMDV)and SVDVcaused similar syndrom in clinical.The assay designed 4 LAMP primers(F3,B3,FIP and BIP)of SVDV VP1 gene sequence,meanwhile the outer primers(F3 and B3)of LAMP primers were used for SVDV PCR detection.Sensitivity,specificity and clinical test were performed.The results showed that sensitivity of the SVDV RT-LAMP was 10 times more higher compared with RT-PCR method.No cross-reactivity with A and O FMDV indicated that RT-LAMP method was specific.Evaluation of the assay with some clinical samples indicated that the SVDV RTLAMP could detect 100%SVDV compared with RT-PCR method.The results indicated that the method of SVDV RT-LAMP was high sensitivity,specificity and rapidity,and was suitable for rapidly detected SVDV in laboratory and clinical.