The development of an automated in situ assay for the detection of human cytomegalovirus in peripheral blood leukocytes

Human cytomegalovirus (HCMV) infections are common in immunosuppressed pati ents, especially transplant recipients and patients with AIDS. The utility of an automated in situ hybridization (ISH) assay for the rapid detection o f HCMV immediate early mRNA was evaluated using cytospin (Shandon Lipshaw, Inc., Pittsburgh, PA) prepared leukocytes from peripheral blood samples. In this study, the detection of HCMV immediate early protein by immunofluores cent antibody staining of the standard shell vial assay was compared to the detection of HCMV immediate early mRNA in peripheral blood leukocytes usin g the automated ISH system. Of 135 specimens tested, eight specimens were p ositive using HCMV ISH compared to seven positive specimens using shell via l assay. Overall, HCMV ISH demonstrated 100% sensitivity and 99% specificit y. Since the HCMV ISH assay requires minimal labor, and can be completed in less than 5 h, this method should be evaluated as a potential replacement for shell vial assay for the diagnosis of HCMV infection. (C) 1999 Elsevier Science B.V. All rights reserved.