Cowley R, Fernandez F, Freemantle W, Rutter D
Division of Pathology, Centre for Applied Microbiology and Research, Salisbury, Wiltshire, United Kingdom.
J Clin Microbiol. 1992 Sep;30(9):2451-5. doi: 10.1128/jcm.30.9.2451-2455.1992.
Enzyme-linked immunosorbent assays (ELISA) for the detection of specific immunoglobulin G (IgG) and IgM antibodies were developed by using purified Coxiella burnetii cells. Variables, including type of microtiter plate, blocking agent, incubation conditions, antigen stability, and substrate type, were examined to achieve optimal ELISA performance. The reliabilities of the assay systems were compared with those of complement fixation (CF) and enhanced immunofluorescence (EIF) tests with 600 human serum samples from defined clinical cases of Q fever, routine samples, and serum specimens from farmers. ELISA and EIF test results agreed in all cases. Dot immunoblotting was also used to test some of these sera and gave a rapid, qualitative result, which agreed with ELISA and EIF test results in all cases. No instances were found in which both ELISA and EIF test results were negative and the CF test results was positive. However approximately 5% of the sera were positive by ELISA and the EIF test while the CF test result was either negative or unreadable because of serum anticomplementary activity. We conclude that dot immunoblotting is a useful screening test, whereas ELISA and the EIF test are both rapid and sensitive tests when used for the serodiagnosis of Q fever and should be considered to be replacements for the CF test.
利用纯化的伯氏考克斯体菌细胞开发了用于检测特异性免疫球蛋白G(IgG)和IgM抗体的酶联免疫吸附测定(ELISA)。对包括微量滴定板类型、封闭剂、孵育条件、抗原稳定性和底物类型在内的变量进行了检测,以实现最佳的ELISA性能。用来自确诊的Q热临床病例、常规样本以及农民血清标本的600份人血清样本,将该检测系统的可靠性与补体结合(CF)试验和增强免疫荧光(EIF)试验的可靠性进行了比较。ELISA和EIF试验结果在所有病例中均一致。斑点免疫印迹法也用于检测其中一些血清,并给出了快速的定性结果,该结果在所有病例中均与ELISA和EIF试验结果一致。未发现ELISA和EIF试验结果均为阴性而CF试验结果为阳性的情况。然而,约5%的血清通过ELISA和EIF试验呈阳性,而CF试验结果因血清抗补体活性而呈阴性或无法读取。我们得出结论,斑点免疫印迹法是一种有用的筛查试验,而ELISA和EIF试验在用于Q热的血清学诊断时都是快速且灵敏的试验,应被视为CF试验的替代方法。