Jahrling P B, Hesse R A, Metzger J F
J Clin Microbiol. 1978 Jul;8(1):54-60. doi: 10.1128/jcm.8.1.54-60.1978.
A radioimmunoassay (RIA) procedure is described for measuring antibodies to alphaviruses in human and other mammalian sera. The test employed protein Abearing Staphylococcus aureus as a solid-phase immunoadsorbent for (3)H-labeled viruses complexed with immunoglobulin G. Using antibodies produced in humans and guinea pigs, the RIA procedure clearly differentiated among antibodies to Venezuelan, western, and eastern equine encephalomyelitis viruses. Sensitivity of the RIA depended on the concentrations of labeled viruses employed. The dilution of serum that effected binding of 50% of the (3)H-labeled virus (determined by probit analysis) was consistently higher than the neutralizing antibody titer determined by a conventional plaque reduction neutralization test using 80% plaque reduction end points. In addition, sera from 73 individuals were screened for seroconversion following live attenuated Venezuelan equine encephalomyelitis virus vaccine (strain TC-83) inoculation, by RIA using a single serum dilution (1:80); results were identical with seroconversions identified by plaque reduction neutralization test. Hyperimmune Venezuelan equine encephalomyelitis virus sera from a number of mammalian species were successfully titrated by RIA; the species tested were human, guinea pig, white rat, rabbit, burro, dog, monkey, sheep, and cotton rat. The protein A-mediated RIA is a rapid, sensitive, specific, and precise serological tool for measuring antibodies to surface antigens of alphaviruses, and should allow the subsequent development of a competitive binding RIA to measure antigenic potency of inactivated alphavirus vaccines.
本文描述了一种放射免疫测定（RIA）方法，用于检测人和其他哺乳动物血清中针对甲病毒的抗体。该测试采用携带蛋白A的金黄色葡萄球菌作为固相免疫吸附剂，用于吸附与免疫球蛋白G结合的³H标记病毒。使用人和豚鼠产生的抗体，RIA方法能够清晰地区分针对委内瑞拉马脑炎病毒、西部马脑炎病毒和东部马脑炎病毒的抗体。RIA的灵敏度取决于所用标记病毒的浓度。通过概率分析确定，使50%的³H标记病毒发生结合的血清稀释度始终高于使用80%蚀斑减少终点的传统蚀斑减少中和试验所确定的中和抗体效价。此外，采用单一血清稀释度（1:80）的RIA方法，对7名个体接种减毒活委内瑞拉马脑炎病毒疫苗（TC-83株）后的血清转化情况进行了筛查；结果与蚀斑减少中和试验所确定的血清转化情况一致。通过RIA成功滴定了多种哺乳动物物种的高免疫委内瑞拉马脑炎病毒血清；所检测的物种包括人、豚鼠、白鼠、兔子、驴、狗、猴子、绵羊和棉鼠。蛋白A介导的RIA是一种快速、灵敏、特异且精确的血清学工具，用于检测针对甲病毒表面抗原的抗体，并且应该能够促进后续竞争性结合RIA的开发，以测量灭活甲病毒疫苗的抗原效力。
