Development of a novel real-time RT-PCR assay with LUX primer for the detection of swine transmissible gastroenteritis virus
Development of a novel real-time RT-PCR assay with LUX primer for the detection of swine transmissible gastroenteritis virus
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DOI：
10.1016/j.jviromet.2004.08.003
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发表时间：
2004-12-01
影响因子：
3.1
通讯作者：
Zhu, DZ
中科院分区：
文献类型：
--
作者：
Chen, R;Huang, WM;Zhu, DZ
实时 RT-PCR 检测基于延伸光 (LUX) 荧光引物和 LightCycle 技术，旨在快速检测传染性胃肠炎病毒 (TGEV)。检测了不同 TGEV 分离株和临床标本的病毒 RNA。为了评估该测定的灵敏度，还开发了一种针对相同 101 bp 序列的基于凝胶的 RT-PCR 方法。通过两种方法检测 TGEV RNA 的连续 10 倍稀释液。尽管实时方法每次反应仅使用 2 μl RNA，但其灵敏度比使用 10 μl RNA 的基于凝胶的方法提高了 10 倍。研究表明，下面报道的 LUX 检测快速、可靠且灵敏，有潜力用作 TGEV 诊断的替代分子方法。 (C) 2004 Elsevier B.V. 保留所有权利。
Real-time RT-PCR assay, based on light upon extension (LUX) fluorogenic primer and LightCycle technology, was developed for rapid detection of transmissible gastroenteritis virus (TGEV). Viral RNA from different TGEV isolates and clinical specimens was detected. To evaluate the sensitivity of the assay, a gel-based RT-PCR method targeted at the same 101 bp sequence was also developed. Serial 10-fold dilutions of TGEV RNA were detected by the two methods. Although the real time method used only 2 mul RNA for each reaction, a 10-fold increase of sensitivity over that of the gel-based method, which used 10 mul RNA was demonstrated. The study indicates that the LUX assay reported below is rapid, reliable and sensitive and it has the potential for use as an alternative molecular method for TGEV diagnosis. (C) 2004 Elsevier B.V. All rights reserved.