Comparison of real-time reverse transcription loop-mediated isothermal amplification and real-time reverse transcription polymerase chain reaction for duck Tembusu virus

详细信息	查看全文 | 推荐本文 |

• 作者：Liping Yan^a ; ¹ ; ^{Liping.yan912@gmail.com} ; Shan Peng^a ; ¹ ; ^{pengshanjiayou2008@163.com} ; Pixi Yan^a ; ¹ ; ^{yanpixi@163.com} ; Jiewen Zhou^a ; ¹ ; ^{jiwen_zhou@sina.com} ; Qiaoyang Teng^a ; ¹ ; ^{tengqy@shvri.ac.cn} ; Guoxin Li^a ; ¹ ; ^{guoxinli@shvri.ac.cn} ; Xuesong Li^a ; ¹ ; ^{lxs116@126.com} ; Zejun Li^a ; ^b ; ^{lizejun@shvri.ac.cn} ; ^{ylp912@126.com}
• 关键词：Reverse transcription loop-mediated isothermal amplification ; Duck Tembusu virus ; Real-time reverse transcription polymerase chain reaction ; Comparison
• 刊名：Journal of Virological Methods
• 出版年：2012
• 期刊代码：180_01660934
• 类别：med
• 出版时间：June, 2012
• 卷：182
• 期：1-2
• 页码：50-55
• 文件大小：831 K

摘要

Duck Tembusu virus (DTMUV) has caused huge losses to the poultry industry in China since the spring of 2010. The development of a rapid, convenient, and reliable method to diagnose this emerging duck infectious disease is critical. In the present study, a real-time reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was compared with the real-time reverse transcription polymerase chain reaction (RT-PCR) for detection of DTMUV. The sensitivity of real-time RT-LAMP was equal to that of the real-time RT-PCR, with a detection limit of 0.01 ELD₅₀ (50%egg lethal dose). The specificity of the real-time RT-LAMP and real-time RT-PCR was confirmed using RNAs and DNAs extracted from related viruses which cause duck infections. The reproducibility of the real-time RT-PCR assay was better than that of the real-time RT-LAMP. Only three results from 96 tissue samples differed between the real-time RT-LAMP and this real-time RT-PCR, confirming the reliability of these methods. This study indicated that the real-time RT-LAMP is simpler, less time-consuming, and more convenient than the real-time RT-PCR. With its high sensitivity, specificity, and convenience, the real-time RT-LAMP is a practical molecular diagnostic method for rapid and quantitative detection of DTMUV infection in a resource-limited setting.