Nielsen S L, Rønholm E, Sørensen I, Andersen H K
J Clin Microbiol. 1986 Dec;24(6):998-1003. doi: 10.1128/jcm.24.6.998-1003.1986.
An antibody capture enzyme-linked immunosorbent assay (ELISA) that uses horseradish-peroxidase-labeled antigen for the detection of immunoglobulin G (IgG) antibodies to cytomegalovirus (CMV) is described. A microtiter plate was coated with anti-human IgG and consecutively incubated with serum specimens, enzyme-labeled CMV antigen made from CMV-infected cell nuclei, and substrate. The CMV IgG antibody content was determined spectrophotometrically and expressed as absorbance. Furthermore, to reveal any nonspecific reactions, all sera were tested against an enzyme-labeled control antigen made from uninfected cell nuclei. The problem with nonspecific reactions was small and was circumvented by the addition of unlabeled control antigen to the conjugates. For epidemiological studies the test was not as sensitive as other serological tests. On the other hand, the IgG antibody capture ELISA was highly sensitive for detecting the serological antibody response in patients with primary and recurrent CMV infections. Thus, one positive serum remained positive at a serum dilution of 1:10(7). The specificity of the test was shown by a blocking experiment and by testing 126 complement fixation-positive sera, of which 97% were positive. There was a rather good correlation between the complement fixation test and the IgG antibody capture ELISA (rs = 0.79, P less than 0.001). The test is especially useful when tests for CMV antibodies of the IgM, IgA, and IgE classes are run by similar antibody capture ELISAs, since the same procedure and conjugate are used.
描述了一种抗体捕获酶联免疫吸附测定（ELISA），该方法使用辣根过氧化物酶标记的抗原检测巨细胞病毒（CMV）的免疫球蛋白G（IgG）抗体。微量滴定板用抗人IgG包被，然后依次与血清标本、由CMV感染细胞核制成的酶标记CMV抗原和底物孵育。通过分光光度法测定CMV IgG抗体含量，并以吸光度表示。此外，为了揭示任何非特异性反应，所有血清都用由未感染细胞核制成的酶标记对照抗原进行检测。非特异性反应问题较小，通过在结合物中加入未标记的对照抗原得以规避。对于流行病学研究，该检测不如其他血清学检测敏感。另一方面，IgG抗体捕获ELISA在检测原发性和复发性CMV感染患者的血清学抗体反应方面高度敏感。因此，一份阳性血清在1:10(7)的血清稀释度下仍保持阳性。通过阻断实验和检测126份补体结合阳性血清（其中97%为阳性）证明了该检测的特异性。补体结合试验与IgG抗体捕获ELISA之间存在相当好的相关性（rs = 0.79，P小于0.001）。当通过类似的抗体捕获ELISA检测IgM、IgA和IgE类别的CMV抗体时，该检测特别有用，因为使用相同的程序和结合物。
