用于检测大肠杆菌O157抗原血清抗体的阻断酶联免疫吸附测定法的开发。
Development of a blocking enzyme-linked immunosorbent assay for detection of serum antibodies to O157 antigen of Escherichia coli.
作者信息
Laegreid W, Hoffman M, Keen J, Elder R, Kwang J
机构信息
U.S. Meat Animal Research Center, USDA Agricultural Research Service, Clay Center, Nebraska 68933, USA.
出版信息
Clin Diagn Lab Immunol. 1998 Mar;5(2):242-6. doi: 10.1128/CDLI.5.2.242-246.1998.
The O157 antigen of Escherichia coli shares structural elements with lipopolysaccharide (LPS) antigens of other bacterial species, notably Brucella abortus and Yersinia enterocolitica 09, a fact that confounds the interpretation of assays for anti-O157 antibodies. To address this problem, a blocking enzyme-linked immunosorbent assay (bELISA) was designed with E. coli O157:H7 LPS as the antigen and a monoclonal antibody specific for E. coli O157, designated 13B3, as the competing antibody. The bELISA had equivalent sensitivity to, and significantly higher specificity than, the indirect ELISA (iELISA), detecting anti-O157 antibodies in sera from cattle experimentally inoculated with O157:H7. Only 13% of sera from naive heifers vaccinated for or experimentally infected with B. abortus had increased anti-O157 bELISA titers, while 61% of anti-O157 iELISA titers were increased. The bELISA is a sensitive and specific method for the detection of serum antibodies resulting from exposure to E. coli O157.
大肠杆菌的O157抗原与其他细菌物种的脂多糖（LPS）抗原具有共同的结构元件，特别是流产布鲁氏菌和小肠结肠炎耶尔森氏菌O9，这一事实混淆了抗O157抗体检测试验的解读。为了解决这个问题，设计了一种阻断酶联免疫吸附测定（bELISA），以大肠杆菌O157:H7 LPS作为抗原，以一种对大肠杆菌O157特异的单克隆抗体（命名为13B3）作为竞争抗体。bELISA对实验接种O157:H7的牛血清中抗O157抗体的检测灵敏度与间接ELISA（iELISA）相当，但特异性显著更高。仅13%接种过流产布鲁氏菌疫苗或实验感染过流产布鲁氏菌的未接触过病原体的小母牛血清的抗O157 bELISA滴度升高，而抗O157 iELISA滴度升高的比例为61%。bELISA是一种检测因接触大肠杆菌O157而产生的血清抗体的灵敏且特异的方法。
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