Foot-and-mouth disease virus: A first inter-laboratory comparison trial to evaluate virus isolation and RT-PCR detection methods
Foot-and-mouth disease virus: A first inter-laboratory comparison trial to evaluate virus isolation and RT-PCR detection methods
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DOI：
10.1016/j.vetmic.2006.06.001
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发表时间：
2006-10-31
影响因子：
3.3
通讯作者：
De Clercq, K.
中科院分区：
文献类型：
--
作者：
Ferris, N. P.;King, D. P.;De Clercq, K.
5个欧洲参考实验室参加了一项评估其用于检测和分离口蹄疫病毒的常规RT-PCR试验和细胞培养的敏感性和特异性的工作。从疑似口蹄疫或猪水疱病（SVD）病例提交的10个水疱上皮细胞中制备了5组相同的20个编码样本。16个样本来自6个口蹄疫病毒阳性上皮，代表4种不同的血清型（O型和A型各2个，Asia 1型和SAT 2型各1个），2个来自抗原ELISA和细胞培养病毒分离（VI）发现阴性的样本，2个来自SVD病毒阳性上皮。一些口蹄疫病毒阳性样本是由三种初始混悬液连续稀释10倍制备的。每个实验室通过一种或多种可用的RT-PCR程序检测样品，并接种常规用于口蹄疫诊断的细胞培养物，试图分离病毒，其特异性通过抗原ELISA证实。每个实验室中使用的最好的RT-PCR检测方法给出了相似的结果，而细胞培养的灵敏度则不同，一个实验室的灵敏度高，两个实验室的灵敏度中等，另外两个实验室的灵敏度低。样品的原型面板似乎适合于这些测试的外部质量保证，但将受益于包含更多的阴性样品和扩展一个或多个口蹄疫阳性样品滴定系列的系列稀释范围。(c) 2006 Elsevier B.V.版权所有
Five European reference laboratories participated in an exercise to evaluate the sensitivity and specificity of their routinely employed RT-PCR tests and cell cultures for the detection and isolation of foot-and-mouth disease (FMD) virus. Five identical sets of 20 coded samples were prepared from 10 vesicular epithelia, which were derived from submissions from suspect cases of FMD or swine vesicular disease (SVD). Sixteen samples were derived from six FMD virus positive epithelia representing four different serotypes (two each of types O and A and one each of types Asia 1 and SAT 2), two from samples which had been found to be negative by antigen ELISA and virus isolation (VI) in cell culture and two from SVD virus positive epithelia. Some of the FMD virus positive samples were prepared from 10-fold serial dilutions of three of the initial suspensions. Each laboratory tested the samples by one or more of its available RT-PCR procedures and inoculated cell cultures that it routinely uses for FMD diagnosis in attempts to isolate virus, the specificity of which was confirmed by antigen ELISA. The best of the RT-PCR assays used in each laboratory gave comparable results while the sensitivity of cell cultures was variable from high in one laboratory, moderate in two and low in two others. This prototype panel of samples would appear suitable for external quality assurance of these tests but would benefit from the inclusion of more negative samples and an extension in the serial dilution range of one or more of the FMD positive sample titration series. (c) 2006 Elsevier B.V. All rights reserved.