Selb B, Weber B
Institut für Medizinische Virologie, Universitätskliniken Frankfurt am Mainz, Germany.
J Virol Methods. 1994 Apr;47(1-2):15-25. doi: 10.1016/0166-0934(94)90062-0.
Immunoglobulin A and G reactivities to human reovirus were investigated comparatively by Western blot and ELISA in a collection of 120 serum samples. Five different reovirus polypeptides of molecular weights of 127, 79, 54 42 and 40 kDa were visualized by polyacrylamide gel electrophoresis. Of 80 anti-reovirus ELISA-IgG positive serum samples, 97.5% and 15.0% reacted with one or more virus-specific bands in the IgG and IgA Western blot respectively. The 127 and 79 kDa proteins reacted with 100% and 96.2% of the IgG Western blot-positive serum samples (n = 78). IgG reactivities to the 40, 54 and 42 kDa proteins were detected in 96.2%, 80.8% and 20.5% of the immunoblot-positive sera. Compared to the Western blot, the ELISA had a specificity of 94.9% for IgG and 96.3% for IgA detection. The sensitivity of specific IgG and IgA detection by ELISA were 96.3% and 100%, respectively. Using the ELISA, a seroepidemiological survey was carried out on 850 sera collected from October 1989 to September 1992. The overall prevalence of reovirus IgG and IgA antibodies were 53.5% and 15.5% respectively. An age dependent increase of the seroprevalence of anti-reovirus IgG was observed. In contrast, the prevalence of IgA antibodies remained constant during childhood. Seroprevalence peak levels were observed in the 9-11, 20-29, 40-49 and > or = 60 years age group. The results of the study suggest that antireovirus IgG antibodies may persist lifelong, whereas IgA antibodies are detectable for a short time and may indicate a recent reovirus infection. The development of a sensitive and specific ELISA assay for the detection of class-specific immunoglobulins permits further investigation of reovirus infections and may contribute to improving the serological diagnosis of recent infection.
通过蛋白质印迹法和酶联免疫吸附测定(ELISA)对120份血清样本中免疫球蛋白A和G与人呼肠孤病毒的反应性进行了比较研究。通过聚丙烯酰胺凝胶电泳观察到分子量分别为127、79、54、42和40 kDa的五种不同呼肠孤病毒多肽。在80份抗呼肠孤病毒ELISA-IgG阳性血清样本中,分别有97.5%和15.0%在IgG和IgA蛋白质印迹法中与一条或多条病毒特异性条带发生反应。127 kDa和79 kDa的蛋白质分别与100%和96.2%的IgG蛋白质印迹法阳性血清样本(n = 78)发生反应。在96.2%、80.8%和20.5%的免疫印迹阳性血清中检测到了针对40 kDa、54 kDa和42 kDa蛋白质的IgG反应性。与蛋白质印迹法相比,ELISA检测IgG的特异性为94.9%,检测IgA的特异性为96.3%。ELISA检测特异性IgG和IgA的灵敏度分别为96.3%和100%。使用ELISA对1989年10月至1992年9月收集的850份血清进行了血清流行病学调查。呼肠孤病毒IgG和IgA抗体的总体流行率分别为53.5%和15.5%。观察到抗呼肠孤病毒IgG血清阳性率随年龄增长而增加。相比之下,IgA抗体的流行率在儿童期保持恒定。在9 - 11岁、20 - 29岁、40 - 49岁和≥60岁年龄组中观察到血清阳性率峰值水平。研究结果表明,抗呼肠孤病毒IgG抗体可能终身存在,而IgA抗体只能在短时间内检测到,并可能表明近期有呼肠孤病毒感染。开发一种用于检测类特异性免疫球蛋白的灵敏且特异的ELISA检测方法,有助于进一步研究呼肠孤病毒感染,并可能有助于改善近期感染的血清学诊断。