Forghani B, Schmidt N J, Lennette E H
Appl Microbiol. 1974 Oct;28(4):661-7. doi: 10.1128/am.28.4.661-667.1974.
A solid phase radioimmunoassay (RIA) was developed for typing Herpesvirus hominis (HVH) strains isolated from clinical materials, and it also proved to be applicable to the direct detection and typing of HVH antigen in human and animal brain tissue. The procedure utilized virus-infected human fetal diploid cells or brain tissue smears in the bottom of 1-dram glass vials, antigen was detected through the use of intermediate HVH antisera produced in rabbits or hamsters and cross-absorbed with the HVH heterotype, and (125)I-labeled anti-species (rabbit or hamster) globulins produced in goats were used for detection of immune complexes. The cross-absorbed HVH antisera could be used at high dilutions in the RIA test, and they reacted with marked type-specificity in the RIA system. Specificity of the test was also improved by determining and using optimal concentrations of intermediate sera and of (125)I-labeled anti-species globulins. Results of typing HVH isolates by the RIA procedure agreed in all instances with those obtained by direct fluorescent antibody staining with cross-absorbed conjugates. The RIA procedure was effective and more sensitive than direct fluorescent antibody for demonstrating and typing HVH antigen directly in smears of infected human brain tissue.
我们开发了一种固相放射免疫测定法(RIA),用于对从临床材料中分离出的人疱疹病毒(HVH)毒株进行分型,并且该方法也被证明适用于直接检测人和动物脑组织中的HVH抗原并进行分型。该方法利用1 dram玻璃瓶底部的病毒感染的人胎儿二倍体细胞或脑组织涂片,通过使用兔或仓鼠产生的并与HVH异型交叉吸收的中间HVH抗血清来检测抗原,并用山羊产生的(125)I标记的抗种属(兔或仓鼠)球蛋白来检测免疫复合物。交叉吸收的HVH抗血清可在RIA试验中以高稀释度使用,并且它们在RIA系统中以明显的型特异性发生反应。通过确定并使用中间血清和(125)I标记的抗种属球蛋白的最佳浓度,也提高了试验的特异性。通过RIA方法对HVH分离株进行分型的结果在所有情况下均与用交叉吸收的结合物进行直接荧光抗体染色所获得的结果一致。RIA方法对于在感染的人脑组织涂片中直接显示和分型HVH抗原是有效的,并且比直接荧光抗体更灵敏。