CAJ | 학술논문

通过对施马伦贝格病毒（Schmallenberg virus，SBV）及其同属各病毒基因序列进行比对分析，设计了针对SBV S基因节段的一组6条特异性引物，经优化建立了SBV RT-LAMP检测方法，可在63℃恒温条件下，于50min内完成目的序列扩增，通过染料法和琼脂糖凝胶电泳法观察结果，其灵敏度可达10TCID50，与荧光RT-PCR方法具有相同的敏感性。特异性试验显示，本方法与同属的沙门达病毒（Shamonda virus）会发生交叉反应，需借助沙门达病毒S基因节段上的一个特异性酶切位点AflII进行酶切，实现两者的鉴别诊断。对103份临床样品的检测结果显示，本方法与荧光RT-PCR方法符合率为100%。本研究建立的RT-LAMP方法快速、简单、灵敏度高，不仅适用于实验室鉴定，还可应用于口岸、野外等的临床大规模监测。
통과대시마륜패격병독（Schmallenberg virus，SBV）급기동속각병독기인서렬진행비대분석，설계료침대SBV S기인절단적일조6조특이성인물，경우화건립료SBV RT-LAMP검측방법，가재63℃항온조건하，우50min내완성목적서렬확증，통과염료법화경지당응효전영법관찰결과，기령민도가체10TCID50，여형광RT-PCR방법구유상동적민감성。특이성시험현시，본방법여동속적사문체병독（Shamonda virus）회발생교차반응，수차조사문체병독S기인절단상적일개특이성매절위점AflII진행매절，실현량자적감별진단。대103빈림상양품적검측결과현시，본방법여형광RT-PCR방법부합솔위100%。본연구건립적RT-LAMP방법쾌속、간단、령민도고，불부괄용우실험실감정，환가응용우구안、야외등적림상대규모감측。
Based on sequence analysis of Schmallenberg virus(SBV),a set of 6 LAMP primers were designed tar-geting the SBV gene S,and an RT-LAMP assay with one step amplification was established after optimization and the assay was finished within 50 minutes at 63℃. The results were observed by color change and agarose gel electropho-resis. The detection limit was 10TCID50,almost as sensitive as the real-time RT-PCR. Considering a cross reaction be-tween SBV and Shamonda virus,a unique AflII restriction site in Shamonda virus was used in the assay and SBV could be distinguished from Shamonda virus. Test of 103 clinical samples showed that the assay was 100%correlated with real-time RT-PCR. The established RT-LAMP was a rapid,simple and sensitive assay,suitable for use both in labora-tory test and clinical field surveillance.