应用环介导逆转录等温扩增技术快速检测新城疫病毒

Reverse Transcriptase Loop-Mediated Isothermal Amplification Technique for Rapid Detection of Newcastle Disease Virus

参考新城疫病毒融合基因(fusion gene,F gene)序列,设计了2对能够特异性地识别F基因上的6个区域的引物,并以此建立了一种简单、快速、有效的环介导逆转录等温扩增(RT-LAMP)方法检测NDV.试验结果表明:该方法用时短(2 h内完成)、特异性好而且灵敏度高,最小检测限可达100 pg;对人工发病样品进行鸡胚分离和RT-LAMP检测,二者的阳性符合率高达93.5%;同时采用镁离子沉淀对检测结果进行可视化处理,方便利用肉眼直接观察.

A simple, rapid and effective reverse transcriptase loop-mediated isothermal amplification（ RT-LAMP） method was developed by using two pairs of specific primers targeting the 6 district sequences of the fusion gene of the newcastle disease virus （NDV）. This RT-LAMP method can rapidly amplified the NDV within 2 h and was highly sensitive and specific. The detection limits was 100 pg by using a serial 10-fold dilution of RNA. Result of detection of clinical samples showed a good correlation between the chicken embryo separation and the RT-LAMP （ coincidence 93.5% ）. The presence of the NDVs could be detected in a simple manner through visual assessment by adding Mg^2＋ into the system to produce a white precipitate.