Eisemann C S, Osterman J V
Am J Trop Med Hyg. 1985 Nov;34(6):1173-8. doi: 10.4269/ajtmh.1985.34.1173.
Hybridoma antibodies (Hab) were prepared against the Karp, Gilliam and Kato strains of Rickettsia tsutsugamushi and were examined for homologous and heterologous reactivity using an indirect immunofluorescence assay. Strain-specific Hab demonstrated homologous IFA titers ranging from 1/320 to 1/1,280 and did not react (less than 1/10) with the heterologous strains. The cross-reactive Hab generally reacted equally with all three strains in the scrub typhus group; however, there were some Hab that reacted with only one of the two heterologous strains tested. The Hab also were examined in enzyme-linked immunosorbent assays with scrub typhus antigens eluted from SDS-polyacrylamide gels. Most Hab reacted with either one or several of the six eluted antigens detected with a polyclonal immune serum. It was also observed that strain-specific and cross-reactive Hab sometimes reacted with the same antigen, suggesting the existence of multiple antigenic determinants in one electrophoretic peak. The data suggest that strain-specific Hab can be used in the indirect immunofluorescence assay to identify isolates of R. tsutsugamushi without the cross-reactions usually observed with polyclonal antisera, and that they are useful probes for detection and analysis of rickettsial antigens.
制备了针对恙虫病东方体卡尔普株、吉利亚姆株和加藤株的杂交瘤抗体(Hab),并使用间接免疫荧光试验检测其同源和异源反应性。菌株特异性Hab的同源免疫荧光抗体效价范围为1/320至1/1280,与异源菌株无反应(小于1/10)。交叉反应性Hab通常与恙虫病组的所有三种菌株反应程度相同;然而,有些Hab仅与所检测的两种异源菌株之一反应。还使用从SDS-聚丙烯酰胺凝胶上洗脱的恙虫病抗原,通过酶联免疫吸附试验检测了Hab。大多数Hab与用多克隆免疫血清检测到的六种洗脱抗原中的一种或几种发生反应。还观察到,菌株特异性和交叉反应性Hab有时会与相同抗原发生反应,这表明在一个电泳峰中存在多个抗原决定簇。数据表明,菌株特异性Hab可用于间接免疫荧光试验,以鉴定恙虫病东方体分离株,而不会出现通常在多克隆抗血清中观察到的交叉反应,并且它们是检测和分析立克次体抗原的有用探针。