Bluetongue virus: European Community inter-laboratory comparison tests to evaluate ELISA and RT-PCR detection methods
Bluetongue virus: European Community inter-laboratory comparison tests to evaluate ELISA and RT-PCR detection methods
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DOI：
10.1016/j.vetmic.2007.11.005
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发表时间：
2008-05-25
影响因子：
3.3
通讯作者：
Oura, C. A. L.
中科院分区：
文献类型：
--
作者：
Batten, C. A.;Bachanek-Bankowska, K.;Oura, C. A. L.
2006年，欧洲共同体国家参考实验室参加了两次实验室间比较测试，以评估其“内部”酶联免疫吸附试验和RT-PCR检测蓝舌病毒抗体和核糖核酸的敏感性和特异性。第一个环试验确定了实验室检测所有24种BTV血清型抗体的能力。第二个环试验包括从实验上感染北欧BTV-8毒株的动物身上采集的抗血清和EDTA血液样本，确定了实验室检测BTV-8抗体和RNA的能力，以及这些检测方法的诊断敏感性。共使用6种C-ELISA、6种实时RT-PCR和3种常规RT-PCR方法。所有C-ELISA都能够检测到目前在欧洲流行的BTV血清型(BTV-1、24、8、9和16)，但一些检测方法对其他血清型的检测不一致，特别是BTV-19。所有的C-ELISA检测牛和羊的BTV-8抗体的时间都是21dpi，而大多数C-ELISA检测牛和羊的抗体是9dpi和8dpi。所有的RT-PCR方法都能检测到BTV-8，但实时检测比常规方法更敏感。大多数实时荧光RT-PCR方法最早在牛和羊体内分别检测到2dpi和3dpi的BTV RNA。这两个环路提供了证据，表明欧共体内的国家参考实验室能够检测BTV抗体和RNA，并提供关于目前可用的检测方法的特异性和敏感性信息。(C)2007 Elsevier B.V.保留所有权利。
European Community national reference laboratories participated in two inter-laboratory comparison tests in 2006 to evaluate the sensitivity and specificity of their 'in-house' ELISA and RT-PCR assays for the detection of bluetongue virus (BTV) antibodies and RNA. The first ring trial determined the ability of laboratories to detect antibodies to all 24 serotypes of BTV. The second ring trial, which included both antisera and EDTA blood samples from animals experimentally infected with the northern European strain of BTV-8, determined the ability of laboratories to detect BTV-8 antibodies and RNA, as well as the diagnostic sensitivity of the assays. A total of six C-ELISAs, six real-time RT-PCR and three conventional RTPCR assays were used. All C-ELISAs were capable of detecting the BTV serotypes currently circulating in Europe (BTV-1, 2 4, 8, 9 and 16), however some assays displayed inconsistencies in the detection of other serotypes, particularly BTV-19. All C-ELISAs detected BTV-8 antibodies in cattle and sheep by 21 dpi, while the majority of assays detected antibodies by 9 dpi in cattle and 8 dpi in sheep. All the RT-PCR assays were able to detect BTV-8, although the real-time assays were more sensitive compared to the conventional assays. The majority of real-time RT-PCR assays detected BTV RNA as early as 2 dpi in cattle and 3 dpi in sheep. These two ring trails provide evidence that national reference laboratories within the EC are capable of detecting BTV antibodies and RNA and provide specificity and sensitivity information on the detection methods currently available. (C) 2007 Elsevier B.V. All rights reserved.