中国实验动物学报2016，Vol.24Issue(3)：293-298,6.DOI:10.3969/j.issn.1005-4847.2016.03.015
仙台病毒RT-LAMP可视化检测方法的建立
Establishment of a visualized detection method of Sendai virus by reverse transcription loop-mediated isothermal amplification
摘要
Abstract
Objective To establish a simple and sensitive detection method of Sendai virus ( SeV ) by reverse transcription loop-mediated isothermal amplification ( RT-LAMP) technique. Methods According to the published Gen-Bank sequences (DQ219803. 1), six pairs of primers were designed targeting the conserved region of SeV. The amplifica-tion products were detected with a LAMP real-time Turbidimeter. (LA-302). Through optimizing the LAMP primers and re-action conditions, a rapid and specific detection method of SeV was established. Meanwhile, the amplified products were colored by fluorescence detection reagent after completion of the reaction, so that the amplification could be visualized and detected by naked eyes. Then, methodological evaluation of the RT-LAMP was tested. Results The method of RT-LAMP showed a highly efficient amplification for SeV viral target gene which was performed at 63℃ for 60 min with the LAMP re-al-time Turbidimeter (LA-302). The detection limit was 2. 1 TCID50, 100 times higher than that of RT-PCR, and no cross-reaction with other RNA and DNA viruses of mice was observed. The results of SeV LAMP reaction was visualized and the tube could be directly observed by naked eyes with the addition of fluorescence detection reagent. The results were consist-ent with the results detected by real-time tubidimeter. 92 clinical samples were detected byRT- LAMP, RT-PCR and indi- rect ELISA, and the coincidence rate was 100%. Conclusions This established SeV RT-LAMP detection method is fast, specific, highly sensitive,easy to perform under simple conditions, and is suitable for rapid detection of Sendai viirus.关键词
仙台病毒/RT-LAMP/实时监控/快速检测Key words
Sendai Virus/RT-LAMP/Real time/Rapid detection分类
生物学引用本文复制引用
周洁,赵丽娟,陶凌云,倪丽菊,高诚,陈洪岩..仙台病毒RT-LAMP可视化检测方法的建立[J].中国实验动物学报,2016,24(3):293-298,6.基金项目
上海市科委科技创新行动计划（11140901000）；上海市科委研发平台专项（15DZ2292400）。 （）
