摘要
目的建立一种检测肠道病毒71型(EV71)快速、敏感的一步逆转录-环介导等温扩增(RT-LAMP)方法。方法针对EV71病毒VP2基因特异性序列的6个区域设计4条LAMP引物,建立RT-LAMP检测方法,并评价其特异性和灵敏度。结果通过GoldView染色和凝胶电泳均能观察到LAMP扩增产物的存在,且与柯萨奇病毒A16型(CA16)无交叉反应发生。所建立的RT-LAMP检测方法灵敏,最低检测限为1.0×102copies/mL。RT-LAMP检测的41份咽拭子标本中有27份出现EV71阳性反应,与荧光定量PCR结果一致。结论 RT-LAMP是一种快速、敏感、特异、准确的方法,适合用于基层医疗机构临床检测。
Objective To establish a reverse transcription loop-mediated isothermal amplification （RT-LAMP） assay for rapid and sensitive detection of enterovirus 71（EV71）. Methods Four primers which recognized 6 distinct regions on the VP2 gene of EV71 were designed for RT-LAMP assay, and the sensitivity and specificity were also evaluated. Results The amplified products were observed with GoldView staining and electrophoresis analysis , and there was no cross reaction with Coxasckie virus A16. The RT-LAMP assay was highly sensitive and had a detection limit of 1.0×10 2 copies/mL. Of 41 swab specimens, 27 were EV71 positive by RT-LAMP assay, which shared 100% identity with fluorescence quantitative PCR. Conclusion RT-LAMP is a rapid, sensitive, specific and accurate method for detection of EV71 in clinical specimens, and especially adapts to primary health care agencies.
出处
《分子诊断与治疗杂志》
2012年第1期26-29,共4页
Journal of Molecular Diagnostics and Therapy
基金
江苏省重点医学人才项目(ky0904)
