Comparison of quantitative cytomegalovirus antigenemia assay with culture methods and correlation with clinical...

Blood samples, obtained predominantly from human immunodeficiency virus-infected patients and solid-organ and bone marrow transplant recipients, were submitted to the clinical laboratory for detection of cytomegalovirus (CMV) and were processed by three methods: conventional culture, centrifugation culture, and CMV antigenemia assay with monoclonal antibodies (Clonab CMV; Biotest Diagnostic Corporation, Denville, N.J.) to CMV antigens. Of 496 blood samples tested, 107 were positive by one or more methods: 56 were positive by conventional culture, 27 were positive by centrifugation culture, and 97 were positive for CMV antigen (Ag) by the antigenemia assay. Forty-seven samples were positive by the CMV antigenemia assay only; in these samples, a mean of 12 Ag-positive cells was detected per 200,000 polymorphonuclear leukocytes examined. In contrast, samples positive by the CMV antigenemia assay and both culture methods had a mean of 193 Ag-positive cells, and samples positive by the CMV antigenemia assay and conventional culture alone had a mean of 157 Ag-positive cells. In the antigenemia assay, paraformaldehyde fixation resulted in superior cell morphology when compared with acetone fixation. Use of immunofluorescence staining reduced sample processing time and the complexity of reagent preparation in comparison with immunoperoxidase staining. Differences in the sensitivities between the immunofluorescence and immunoperoxidase staining techniques for detection of antigenemia were minor, with discrepant samples showing only one or two Ag-positive cells. Clinical disease was generally associated with high-level antigenemia, but exceptions were noted. The CMV antigenemia test is a rapid, quantitative assay that greatly facilitated the rapid diagnosis of CMV infection. However, quantitation of antigenemia is labor-intensive, requires processing of samples soon after collection, and does not always correlate with clinical disease in the individual patient.