The development of an accelerated reverse-transcription loop mediated isothermal amplification for the serotype specific detection of bluetongue virus 8 in clinical samples
The development of an accelerated reverse-transcription loop mediated isothermal amplification for the serotype specific detection of bluetongue virus 8 in clinical samples
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DOI：
10.1016/j.jviromet.2014.03.004
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发表时间：
2014-06-15
影响因子：
3.1
通讯作者：
Welsh, Michael D.
中科院分区：
文献类型：
--
作者：
Mulholland, Catherine;Hoffmann, Bernd;Welsh, Michael D.
2006年，荷兰首次发现蓝舌病病毒血清型8（BTV 8），导致绵羊和牛的大流行，并迅速蔓延到邻国比利时，德国以及法国和英国。这导致了严重的动物健康和福利问题，并给这些国家的农业食品工业造成了巨大的经济损失。鉴于BTV感染的早期诊断“现场”对其后续管理和控制非常有用，本研究旨在设计一种新型，灵敏和快速的核酸诊断测试用于BTV 8的病毒型特异性检测，该测试可以在不使用先进的实验室支持和设备的情况下使用。用于检测BTV 8的引物基于病毒的基因组片段2，即VP 2基因。使用一组完整的BTV参考株和临床样本对该检测进行评估。使用荧光检测试剂观察到阳性扩增。RT-LAMP检测的灵敏度为102个拷贝的RNA。该试验未扩增密切相关的环状病毒EHDV。这种新型的RT-LAMP提供了一种灵敏、特异和快速的检测BTV 8的方法。这种方法价格低廉，易于使用，有可能在“现场”的“笔侧”环境中使用，或在发展中国家的较小的、设备较差的实验室中使用。(C)2014爱思唯尔有限公司版权所有。
In 2006 bluetongue virus serotype 8 (BTV 8) was identified for the first time in the Netherlands causing a major epidemic in sheep and cattle that quickly spread to neighbouring Belgium, Germany and beyond to France and the UK. This resulted in severe animal health and welfare problems as well as substantial economic losses to the agrifood industries of these countries. Given that the early diagnosis of BTV infection 'in-the-field' is extremely useful to its subsequent management and control, this study was established to design a novel, sensitive and rapid nucleic acid diagnostic test for the serotype-specific detection of BTV 8, which could be used without the use of advanced laboratory support and equipment. Primers for the detection of BTV 8 were based on genome segment 2 of the virus, the VP2 gene. The assay was assessed using a full panel of BTV reference strains and clinical samples. Positive amplification was observed using a fluorescent detection reagent. The sensitivity of the RT-LAMP assay was 102 copies of RNA. The assay did not amplify the closely related orbivirus EHDV. This novel RT-LAMP offers a sensitive, specific and rapid method of detecting BTV 8. The approach is inexpensive and easy to use and could potentially be used in a 'pen-side' setting 'in the field' or by smaller less well-equipped laboratories in developing countries. (C) 2014 Elsevier B.V. All rights reserved.