Use of Sindbis/Eastern Equine Encephalitis Chimeric Viruses in Plaque Reduction Neutralization Tests for Arboviral Disease Diagnostics
Use of Sindbis/Eastern Equine Encephalitis Chimeric Viruses in Plaque Reduction Neutralization Tests for Arboviral Disease Diagnostics
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DOI：
10.1128/cvi.05129-11
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发表时间：
2011-09-01
影响因子：
--
通讯作者：
Weaver, S. C.
中科院分区：
文献类型：
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作者：
Johnson, B. W.;Kosoy, O.;Weaver, S. C.
东部马脑炎病毒（EEEV）是一种高毒性的蚊媒甲病毒，在北美东部、加勒比海、墨西哥以及整个中美洲和南美洲的人类和马中引起严重且通常致命的神经系统疾病。EEEV感染通过抗EEEV特异性IgM检测进行血清学诊断，并通过空斑减少中和试验（PRNT）进行确认，该试验对甲病毒具有高度特异性。在PRNT程序中使用活病毒，目前需要生物安全3级遏制设施，并在EEEV情况下选择代理安全。这些要求限制了公共卫生实验室进行PRNT的能力。辛德毕斯病毒（SINV）/EEEV重组构建体已经被工程化以减毒形式表达来自2种野生型EEEV毒株的免疫原性结构蛋白。这些SINV/EEEV未被归类为选择因子，使用人、马和鼠血清在PRNT中作为替代诊断试剂进行了评估。结果表明，嵌合病毒表现出与野生型EEEV相当的特异性，只有轻微的灵敏度降低。考虑到它们在增加安全性和降低监管要求方面的益处，这些嵌合病毒在整个美洲的诊断实验室中应该非常有用。
Eastern equine encephalitis virus (EEEV) is a highly virulent, mosquito-borne alphavirus that causes severe and often fatal neurological disease in humans and horses in eastern North American, the Caribbean, and Mexico and throughout Central and South America. EEEV infection is diagnosed serologically by anti-EEEV-specific IgM detection, with confirmation by the plaque reduction neutralization test (PRNT), which is highly specific for alphaviruses. Live virus is used in the PRNT procedure, which currently requires biosafety level 3 containment facilities and select agent security in the case of EEEV. These requirements restrict the ability of public health laboratories to conduct PRNTs. Sindbis virus (SINV)/EEEV recombinant constructs have been engineered to express the immunogenic structural proteins from 2 wild-type EEEV strains in an attenuated form. These SINV/EEEVs, which are not classified as select agents, were evaluated as alternative diagnostic reagents in a PRNT using human, equine, and murine sera. The results indicate that the chimeric viruses exhibit specificity comparable to that of wild-type EEEV, with only a slight reduction in sensitivity. Considering their benefits in increased safety and reduced regulatory requirements, these chimeric viruses should be highly useful in diagnostic laboratories throughout the Americas.