Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of African Horse Sickness Virus.
Development of a Novel Reverse Transcription Loop-Mediated Isothermal Amplification Assay for the Rapid Detection of African Horse Sickness Virus.
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DOI：
10.1111/tbed.12549
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发表时间：
2017-10
影响因子：
4.3
通讯作者：
Castillo-Olivares J
中科院分区：
文献类型：
--
作者：
Fowler VL;Howson ELA;Flannery J;Romito M;Lubisi A;Agüero M;Mertens P;Batten CA;Warren HR;Castillo-Olivares J
非洲马瘟（AHS）是由非洲马瘟病毒（AHSV）引起的一种马科动物疾病，由库蠓传播。AHS是撒哈拉以南非洲的地方病，但在过去的一个世纪里，北方非洲国家、伊比利亚和阿拉伯半岛、中东和印度次大陆都记录了具有重大经济意义和死亡率升高的疫情。有效的控制结合了早期预警系统的应用，准确的实验室诊断和报告，动物移动限制，适当的疫苗接种和监测计划，以及有效的兽医服务对所有这些措施的协调。传统的逆转录酶（RT）PCR（RT-PCR）和真实的实时RT-PCR（rRT-PCR）检测提高了诊断AHS的灵敏度和快速性，因此世界动物卫生组织（OIE）将这些方法作为推荐的检测方法。然而，目前这些检测仅在实验室环境中进行，因此，AHS现场诊断的发展将提高控制政策的快速实施。环介导等温扩增（LAMP）是一种等温、自动循环、链置换核酸扩增技术，可在现场进行。LAMP检测是有吸引力的分子检测，因为它们使用简单、快速、便携，并且在rRT-PCR范围内具有灵敏度和特异性。本研究描述了用于检测AHSV的新型RT-LAMP试验的开发。当考虑rRT-PCR临界值C T > 36时，AHSV RT-LAMP试验的分析灵敏度为96.1%，或当不应用rRT-PCR临界值时为91.3%。诊断敏感性和特异性均为100%。该测定提供了用于现场的快速且低成本的AHS诊断。
African horse sickness (AHS) is a disease of equids caused by African Horse Sickness Virus (AHSV) and is transmitted by Culicoides midges. AHS is endemic in sub‐Saharan Africa, but during the past century, outbreaks of significant economic importance and elevated mortality have been recorded in Northern African countries, the Iberian and Arabian Peninsula, the Middle East and the Indian subcontinent. Effective control combines the application of early warning systems, accurate laboratory diagnosis and reporting, animal movement restrictions, suitable vaccination and surveillance programs, and the coordination of all these measures by efficient veterinary services. Conventional reverse‐transcriptase (RT) PCR (RT‐PCR) and real‐time RT‐PCR (rRT‐PCR) assays have improved the sensitivity and rapidity of diagnosing AHS, resulting in the adoption of these methods as recommended tests by the World Organisation for Animal Health (OIE). However, currently these assays are only performed within laboratory settings; therefore, the development of field diagnostics for AHS would improve the fast implementation of control policies. Loop‐mediated isothermal amplification (LAMP) is an isothermal, autocycling, strand‐displacement nucleic acid amplification technique which can be performed in the field. LAMP assays are attractive molecular assays because they are simple to use, rapid, portable and have sensitivity and specificity within the range of rRT‐PCR. This study describes the development of a novel RT‐LAMP assay for the detection of AHSV. The AHSV RT‐LAMP assay has an analytical sensitivity of 96.1% when considering an rRT‐PCR cut‐off value of C T > 36, or 91.3% when no rRT‐PCR cut‐off is applied. Diagnostic sensitivity and specificity were 100%. This assay provides for a rapid and low cost AHS diagnostic for use in the field.
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影响因子：
3.7
作者：
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通讯作者：
LAEGREID, WW
影响因子：
3.8
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影响因子：
2.4
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影响因子：
3.1
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Mondal, Bimalendu
影响因子：
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作者：
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通讯作者：
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