Comparison of real-time reverse transcription loop-mediated isothermal amplification and real-time reverse transcription polymerase chain reaction for duck Tembusu virus
Comparison of real-time reverse transcription loop-mediated isothermal amplification and real-time reverse transcription polymerase chain reaction for duck Tembusu virus
复制标题
鸭天布苏病毒实时逆转录环介导等温扩增与实时逆转录聚合酶链反应的比较
DOI：
10.1016/j.jviromet.2012.03.007
复制
发表时间：
2012-06-01
影响因子：
3.1
通讯作者：
Li, Zejun
中科院分区：
文献类型：
--
作者：
Yan, Liping;Peng, Shan;Li, Zejun
自2010年春季以来，鸭坦布苏病毒（DTMUV）给中国家禽业造成了巨大损失。开发一种快速、方便、可靠的方法来诊断这种新出现的鸭传染病是至关重要的。本研究将实时逆转录环介导的等温扩增（RI-LAMP）法与实时逆转录聚合酶链反应（RT-PCR）法检测DTMUV进行了比较。real-time RT-LAMP的灵敏度与real-time RT-PCR相当，检测限为0.01 ELD50（50%卵致死剂量）。利用从引起鸭感染的相关病毒中提取的rna和dna，验证了实时RT-LAMP和实时RT-PCR的特异性。实时RT-PCR法的重现性优于实时RT-LAMP法。在96个组织样本中，实时RI-LAMP和实时RT-PCR之间只有3个结果不同，证实了这些方法的可靠性。本研究表明，real-time RI-LAMP比real-time RT-PCR更简单、更省时、更方便。实时RT-LAMP具有高灵敏度、特异性和便捷性，是一种在资源有限的环境下快速、定量检测DTMUV感染的实用分子诊断方法。(C) 2012 Elsevier B.V.版权所有
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 (RI-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 RI-LAMP and this real-time RT-PCR, confirming the reliability of these methods. This study indicated that the real-time RI-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. (C) 2012 Elsevier B.V. All rights reserved.