Assessment of plaque assay methods for alphaviruses
Assessment of plaque assay methods for alphaviruses
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DOI：
10.1016/j.jviromet.2012.09.026
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发表时间：
2013-01-01
影响因子：
3.1
通讯作者：
Halsey, Eric S.
中科院分区：
文献类型：
--
作者：
Juarez, Diana;Long, Kanya C.;Halsey, Eric S.
甲型病毒属的病毒是影响世界各地人类健康的许多虫媒病毒疾病的罪魁祸首。急性甲型病毒感染的确认是基于病毒分离、病毒RNA鉴定或急性和恢复期样本之间抗体效价增加四倍或更多。在恢复期，甲型病毒抗体的特异性可通过空斑减少中和试验来证实。为了确定识别甲型病毒和中和抗体的最佳方法，评价了使用0.4%琼脂糖单层细胞覆盖的标准固体方法和使用0.6%羧甲基纤维素(CMC)覆盖的细胞悬浮覆盖的半固体方法。选择Mayaro病毒、Una病毒、委内瑞拉马脑炎病毒(VEEV)和西方马脑炎病毒(Weev)进行两种方法的检测。结果表明，固体法的灵敏度始终高于半固体法。同时，一种使用0.6%CMC覆盖在细胞单层上的“半固体变异法”被用于病毒滴定。该方法对VEEV的灵敏度与固相法相同，而对WeEV滴定的灵敏度更高。斑块检测条件的改变对结果有很大影响，因此需要对每一种新检测方法的性能进行评估。爱思唯尔出版公司(Elsevier B.V.)
Viruses from the Alphavirus genus are responsible for numerous arboviral diseases impacting human health throughout the world. Confirmation of acute alphavirus infection is based on viral isolation, identification of viral RNA, or a fourfold or greater increase in antibody titers between acute and convalescent samples. In convalescence, the specificity of antibodies to an alphavirus may be confirmed by plaque reduction neutralization test. To identify the best method for alphavirus and neutralizing antibody recognition, the standard solid method using a cell monolayer overlay with 0.4% agarose and the semisolid method using a cell suspension overlay with 0.6% carboxymethyl cellulose (CMC) overlay were evaluated. Mayaro virus, Una virus, Venezuelan equine encephalitis virus (VEEV), and Western equine encephalitis virus (WEEV) were selected to be tested by both methods. The results indicate that the solid method showed consistently greater sensitivity than the semisolid method. Also, a "semisolid-variant method" using a 0.6% CMC overlay on a cell monolayer was assayed for virus titration. This method provided the same sensitivity as the solid method for VEEV and also had greater sensitivity for WEEV titration. Modifications in plaque assay conditions affect significantly results and therefore evaluation of the performance of each new assay is needed. Published by Elsevier B.V.