Scott T W, Olson J G
Am J Trop Med Hyg. 1986 May;35(3):611-8. doi: 10.4269/ajtmh.1986.35.611.
An enzyme immunoassay (EIA) was evaluated for its efficacy at detecting eastern equine encephalomyelitis (EEE) virus in avian blood and brain specimens. Preliminary analysis of blood from experimentally infected house sparrows and naturally infected whooping cranes showed that EEE antigen could be detected with the EIA. Polyclonal mouse antibodies were selected for antigen capture, and rabbit antibodies were selected for antigen detection. Overnight antigen incubation increased sensitivity. The lower limit of EEE antigen detection was 10(3.5) TCID50/ml for a stock of virus. Sensitivity was 10% (2/20) for antigen detection in the blood of chicks inoculated with EEE virus less than 24 hr earlier. At 24 and 48 hr after infection, sensitivity was 100% (10/10). Sensitivity and specificity of antigen detection were excellent (100% for both) in house sparrows experimentally inoculated with EEE, Highlands J (HJ), western equine encephalomyelitis (WEE), or St. Louis encephalitis (SLE) virus and bled at 24 hr intervals. Cross-reactivity was observed, however, with high concentrations (10(5.5) TCID50/ml) of HJ virus. EEE antigen was detected in avian blood by the EIA after infectious virus had declined to undetectable levels. The EIA is a useful alternative to virus isolation in cell culture for diagnosis or detection of EEE virus infections in birds. The test has the advantages of being simple, rapid, and capable of detecting antigen in the absence of infectious virus.
对一种酶免疫测定法（EIA）检测禽血和脑标本中东马脑炎（EEE）病毒的效果进行了评估。对实验感染的家麻雀和自然感染的美洲鹤血液的初步分析表明，EIA能够检测到EEE抗原。选择多克隆小鼠抗体进行抗原捕获，选择兔抗体进行抗原检测。过夜抗原孵育可提高灵敏度。对于一批病毒，EEE抗原检测的下限为10(3.5) TCID50/ml。对于接种EEE病毒不到24小时的雏鸡血液中的抗原检测，灵敏度为10%（2/20）。感染后24小时和48小时，灵敏度为100%（10/10）。在实验接种EEE、高地J（HJ）、西马脑炎（WEE）或圣路易斯脑炎（SLE）病毒并每隔24小时采血的家麻雀中，抗原检测的灵敏度和特异性极佳（均为100%）。然而，观察到与高浓度（10(5.5) TCID50/ml）的HJ病毒存在交叉反应。在感染性病毒降至无法检测的水平后，EIA仍能在禽血中检测到EEE抗原。EIA是细胞培养病毒分离的一种有用替代方法，可用于诊断或检测鸟类的EEE病毒感染。该检测方法具有简单、快速的优点，并且能够在无感染性病毒的情况下检测抗原。
