Detection of Japanese Encephalitis Virus Antigens by the Sandwich ELISA in Infected Cell Culture Fluid and Cell Homogenates
Detection of Japanese Encephalitis Virus Antigens by the Sandwich ELISA in Infected Cell Culture Fluid and Cell Homogenates
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夹心 ELISA 检测感染细胞培养液和细胞匀浆中的日本脑炎病毒抗原
DOI：
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发表时间：
1989
期刊：
影响因子：
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通讯作者：
K. Bundo
中科院分区：
文献类型：
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作者：
K. Bundo
采用夹心ELISA法检测白纹伊蚊、克隆C6/36和BHK21细胞感染不同浓度乙脑病毒后培养液和匀浆中的乙脑病毒抗原。在这两个细胞系统中，当感染病毒滴度升至 107 PFU/ml 时，感染液体中的 ELISA 抗原即可检测到，并且 ELISA 滴度在达到平台后仍保持在相似的水平。乙脑病毒在高输入感染复数下在两个细胞系统中的生长曲线实验表明，在感染的BHK21细胞培养液和匀浆以及感染的C6/36细胞培养液中，病毒感染性、血凝活性（HA）和ELISA抗原的滴度几乎平行上升。然而，在感染的C6/36细胞匀浆中，HA滴度保持在不可检测的水平，而感染性和ELISA滴度则上升。对受感染的 BHK21 和 C6/36 细胞培养液和匀浆进行的密度梯度沉降分析表明，在缓慢沉降的轻级分（酒石酸钾中密度为 1.12 g/cc）中检测到大量病毒抗原，而在与病毒感染性相关的快速沉降的重级分（密度 1.16 g/cc）中检测到较少量的病毒抗原。乙脑病毒ELISA抗原检测采用现场采集的三带喙库蚊匀浆接种的C6/36细胞培养液，并保持在28℃。接种后 2 至 5 天，所有 9 个池中的乙脑 - ELISA 抗原均变得可检测，最终通过既定方法进行乙脑病毒分离，结果呈阳性，而所有 11 个池，通过乙脑病毒分离结果呈阴性，除了单个样本显示短暂和临界水平（1 个单位）的反应外，没有产生可检测水平的乙脑 - ELISA 抗原。总共 256 份来自 C6/36 细胞培养物的培养液，接种了现场捕获的 Cx。 tritaeniorhynchus 于 1978 年至 1989 年 4 月 27 日收稿。长崎大学热带医学研究所投稿号 2248。
Sandwich ELISA was applied to detect Japanese encephalitis (JE) virus antigens in culture fluid and homogenate of Aedes albopictus, clone C6/36, and BHK21 cells daily after infection with various concentrations of JE virus. In both cell systems, the ELISA antigen in the infected fluid became detectable when infective virus titer rose up to 107 PFU/ml, and the ELISA titer remained at similar levels after reaching its plateau. Growth curve experiment of JE virus in both cell systems at high input multiplicity of infection showed that the titers of virus infectivity, hemagglutinating activity (HA), and ELISA antigen rose up almost in parallel in the infected BHK21 cell culture fluid and homogenate as well as in the infected C6/36 cell culture fluid. In the infected C6/36 cell homogenate, however, the HA titer remained at undetectable level while infectivity and ELISA titers rose up. Density gradient sedimentation analysis on the infected BHK21 and C6/36 cell culture fluids and homogenates showed that larger amounts of the virus antigen were detected in the slowly sedimenting light fraction (density 1.12 g/cc in potassium tartrate), with less amounts in the rapidly sedimenting heavy fraction associated with virus infectivity (densty 1.16 g/cc). The detection of JE virus ELISA antigen was applied to the culture fluid of C6/36 cells inoculated with field-caugth Culex tritaeniorhynchus mosquito homogenates and maintained at 28•Ž. The JE-ELISA antigen became detectable between 2 to 5 days after the inoculation for all the 9 pools which eventually turned out to be positive with JE virus isolation by the established method, while all the 11 pools which turned out to be negative with JE viurus isolation did not produce detectable levels of JE-ELISA antigen except a single specimen showing transient and borderline level (1 unit) of the reaction. A total of 256 culture fluids from C6/36 cell cultures inoculated with field-caught Cx. tritaeniorhynchus between 1978 to R eceived for Publication, April 27, 1989. Contribution No. 2248 from the Institute of Tropical Medicine, Nagasaki University.
通过酶免疫分析 (EIA) 检测蚊子池中的东部马脑脊髓炎病毒和高地 J 病毒抗原。
DOI：
10.4269/ajtmh.1984.33.965
发表时间：
1984
期刊：
The American journal of tropical medicine and hygiene
影响因子：
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作者：
Hildreth,SW;Beaty,BJ
通讯作者：
Beaty,BJ
通过酶免疫分析 (EIA) 检测蚊子池中的东部马脑脊髓炎病毒和高地 J 病毒抗原。
DOI：
10.4269/ajtmh.1984.33.973
发表时间：
1984
期刊：
The American journal of tropical medicine and hygiene
影响因子：
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作者：
Hildreth,SW;Beaty,BJ;Maxfield,HK;Gilfillan,RF;Rosenau,BJ
通讯作者：
Rosenau,BJ