The Digene Hybrid Capture System (DHCS) is a solution hybridization antibody capture assay for the chemiluminescent detection and quantitation of cytomegalovirus (CMV) DNA in leukocytes. This assay was compared with the CMV antigenemia assay and shell vial and tube cultures for the detection of CMV in 234 blood specimens from 72 patients with human immunodeficiency virus. Intra- and interrun precision of the DHCS assay gave coefficients of variation of 17.8 and 16.3%, respectively. The correlation coefficient for the quantitative results obtained by the DHCS assay and the antigenemia assay was 0.911 (95% confidence interval, 0.885 to 0.930). Agreement between the DHCS assay and the other three assays ranged from 83 to 86%. The DHCS assay detected 71, 87, and 84% of specimens that were positive by antigenemia, shell vial cultures, and tube culture, respectively. A total of 92% of specimens that were positive by the DHCS assay were also positive by at least one of the other assays. Evaluation of the usefulness of quantitation of CMV DNA by using the DHCS assay and its correlation with clinical disease demonstrated that, with some exceptions, patients with clinical CMV disease tended to have high levels of DNA whereas asymptomatic patients tended to have low or undetectable levels. Overall, the DHCS assay provided a rapid, quantitative, and objective measure of CMV activity in leukocytes, but results did not always correlate with clinical disease.