Viboud G I, Binsztein N, Svennerholm A M
Department of Medical Microbiology and Immunology, University of Göteborg, Sweden.
J Clin Microbiol. 1993 Mar;31(3):558-64. doi: 10.1128/jcm.31.3.558-564.1993.
Monoclonal antibodies (MAbs) against five putative colonization factors (PCFs), i.e., colonization factor antigen (CFA)/III, coli surface antigen (CS)7 and CS17, PCFO159, and PCFO166 of enterotoxigenic Escherichia coli (ETEC), were produced. Hybridomas (one each) producing specific antibodies against the respective PCFs were selected. All the MAbs reacted with the corresponding fimbriae but not with CFA/I, CFA/II, or CFA/IV or the heterologous PCFs in bacterial agglutination and enzyme-linked immunosorbent assays (ELISAs). In immunoelectron microscopy these MAbs bound along the fimbriae, and they also reacted with the corresponding subunits in immunoblots. The five MAbs were used to evaluate the prevalence of CFA/III, CS7, CS17, PCFO159, and PCFO166 in ETEC strains isolated from children with diarrhea in Argentina. One hundred five ETEC isolates negative for CFA/I, CFA/II, and CFA/IV were tested in slide agglutination or in a dot blot test for spontaneously agglutinating strains; positive results were confirmed by inhibition ELISAs. It was found that 27% of the CFA-negative ETEC strains carried one of the PCFs. The sensitivity of slide agglutination with these MAbs was similar to that with specific polyclonal antisera; however, the specificity was higher. PCFO166 was found in 9.5% of the strains tested, mainly in ETEC of serogroup O78 producing heat-stable toxin alone. CS17 and CS7 were identified in 6.7 and 5.7%, respectively, of strains producing heat-labile toxin only, most of which belonged to serogroup O114. PCFO159 was found in 3.8% of the isolates tested, whereas CFA/III was detected in only one ETEC strain.
制备了针对产肠毒素大肠杆菌（ETEC）的五种假定定植因子（PCF）的单克隆抗体（MAb），即定植因子抗原（CFA）/III、大肠杆菌表面抗原（CS）7和CS17、PCFO159和PCFO166。筛选出产生针对各自PCF的特异性抗体的杂交瘤（每种一个）。在细菌凝集试验和酶联免疫吸附测定（ELISA）中，所有单克隆抗体均与相应菌毛发生反应，但不与CFA/I、CFA/II或CFA/IV或异源PCF发生反应。在免疫电子显微镜下，这些单克隆抗体沿菌毛结合，并且它们在免疫印迹中也与相应的亚基发生反应。使用这五种单克隆抗体评估从阿根廷腹泻儿童中分离出的ETEC菌株中CFA/III、CS7、CS17、PCFO159和PCFO166的流行情况。对105株CFA/I、CFA/II和CFA/IV阴性的ETEC分离株进行玻片凝集试验或针对自发凝集菌株的斑点印迹试验检测；阳性结果通过抑制ELISA进行确认。发现27%的CFA阴性ETEC菌株携带其中一种PCF。使用这些单克隆抗体进行玻片凝集试验的敏感性与使用特异性多克隆抗血清的敏感性相似；然而，特异性更高。在所检测的菌株中，9.5%的菌株中发现了PCFO166，主要存在于仅产生热稳定毒素的O78血清型ETEC中。仅产生热不稳定毒素的菌株中，分别有6.7%和5.7%的菌株鉴定出CS17和CS7，其中大多数属于O114血清型。在所检测的分离株中，3.8%的菌株中发现了PCFO159，而仅在一株ETEC菌株中检测到CFA/III。
