CAJ | 학술논문

[目的]建立一种快速、敏感、特异的检测牛病毒性腹泻病毒（BVDV）反转录环介导等温扩增（RT-LAMP）方法，为诊断与监测BVDV 提供准确可靠工具。[方法]根据GenBank公布的BVDV序列，在其保守序列区域设计了一套 LAMP引物，优化了反应时间与反应温度，建立了检测 BVDV的 RT-LAMP方法。[结果]该方法能在56℃等温条件下40分钟完成扩增，扩增结果可通过凝胶电泳梯形带判定。该方法具有良好的特异性和敏感性，最低能检测到3.74×100 copies/μl的病毒 RNA，而且与其他病毒无交叉反应。[结论]该方法能用于牛病毒性腹泻病毒的诊断与监测。
[목적]건립일충쾌속、민감、특이적검측우병독성복사병독（BVDV）반전록배개도등온확증（RT-LAMP）방법，위진단여감측BVDV 제공준학가고공구。[방법]근거GenBank공포적BVDV서렬，재기보수서렬구역설계료일투 LAMP인물，우화료반응시간여반응온도，건립료검측 BVDV적 RT-LAMP방법。[결과]해방법능재56℃등온조건하40분종완성확증，확증결과가통과응효전영제형대판정。해방법구유량호적특이성화민감성，최저능검측도3.74×100 copies/μl적병독 RNA，이차여기타병독무교차반응。[결론]해방법능용우우병독성복사병독적진단여감측。
Objective] This study aimed to develop a reverse transcription loop-medi-ated isothermal amplification (RT-LAMP) method for detecting BVDV. [Method] Since gp48 gene of BVDV is among the most conserved regions, a set of four primers was designed to amplify six target sequences at the gp48 gene region for the RT-LAMP assay. The optimization of the RT-LAMP reaction was performed by evaluat-ing reaction temperature and reaction time. [Result] The RT-LAMP aasay was suc-cessful y conducted at 56 ℃ within 40 min under isothermal conditions, and the re-sults could be detected as ladder-like bands using agarose gel electrophoresis. The RT-LAMP assay is highly sensitive and able to detect 3.74 ×100 copies/μl of BVDV RNA, as no cross-reaction was observed with other viruses. [Conclusion] Overal , the newly established RT-LAMP assay indicates the potential application in both clinical diagnosis and field surveil ance of BVDV.