Rapid and sensitive detection of African horse sickness virus by real-time PCR
Rapid and sensitive detection of African horse sickness virus by real-time PCR
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DOI：
10.1016/j.rvsc.2008.07.015
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发表时间：
2009-04-01
影响因子：
2.4
通讯作者：
Sanchez-Vizcaino, J. M.
中科院分区：
文献类型：
--
作者：
Fernandez-Pinero, J.;Fernandez-Pacheco, P.;Sanchez-Vizcaino, J. M.
一种高度敏感和特异性的TaqMan-MGB实时RT-PCR检测方法已被开发和标准化，用于检测非洲马瘟病毒（AHSV）。在基因组区段7的高度保守区域内选择对AHSV特异性的引物和MGB探针。通过检测来自所有9种血清型的12株AHSV分离株，验证了该诊断方法的稳健性和普遍适用性。根据病毒血清型，分析灵敏度范围为0.001 - 0.15 TCID 50/反应。实时PCR性能进行了初步评估，通过分析一组现场马样本。在没有实时PCR仪器的实验室中，使用相同的引物对来验证常规RT-PCR作为负担得起的、有用的和简单的替代方法。这两项技术为改善非洲马病（AHS）的分子诊断提供了新的工具。(C)2008爱思唯尔有限公司保留所有权利。
A highly sensitive and specific TaqMan-MGB real-time RT-PCR assay has been developed and standardised for the detection of African horse sickness virus (AHSV). Primers and MGB probe specific for AHSV were selected within a highly conserved region of genome segment 7. The robustness and general application of the diagnostic method were verified by the detection of 12 AHSV isolates from all of the nine serotypes. The analytical sensitivity ranged from 0.001 to 0.15 TCID50 per reaction, depending on the viral serotype. Real-time PCR performance was preliminarily assessed by analysing a panel of field equine samples. The same primer pair was used to standardise a conventional RT-PCR as an affordable, useful and simple alternative method in laboratories without access to real-time PCR instruments. The two techniques present novel tools to improve the molecular diagnosis of African horse sickness (AHS). (C) 2008 Elsevier Ltd. All rights reserved.