Cell Culture-PCR Technique for Detection of Infectious Cytomegalovirus in Peripheral Blood

WedeterminedtheefficacyofPCRforthedirectdetectionofinfectiouscytomegalovirus(CMV)inspecimeninoculated MRC-5 tube cultures (cell culture-PCR [CC-PCR]). Parallel testing was performed by a shell vial assay-indirect immunofluorescence assay (SVA-IFA) and isolation (conventional MRC tube cultures [TCCPE]. The sensitivity, specificity, and positive and negative predictive values for CC-PCR, SVA-IFA, and TC-CPE were 81, 99, 94, and 94%, 86, 100, 100, and 96%, and 77, 100, 100, and 93%, respectively. Future application of CC-PCR may be directed toward its use as a screening tool for cytomegalovirus genotypic variants. The detection of productive cytomegalovirus (CMV) infection in blood leukocytes serves as a marker and has prognostic value for the development of CMV disease in immunocompromised patients. In an effort to improve test sensitivity, direct detection of CMV in the blood leukocyte fraction by PCR technology has been performed. However, this genomic detection methodology does not always correlate with CMV-related disease (4, 10). Detection of CMV in plasma or serum by PCR and detection of antigen in broncheoalveolar lavage or by serologic methods have also been performed. These procedures retain the limitations of low correlation with cell culture (11, 14) or clinical (CMV) disease (1, 5). In an attempt to utilize the sensitivity of CMV DNA PCR but preserve clinical relevance through the detection of infectiousvirus,anovelcellculture-PCR(CC-PCR)assaywasevaluated in our laboratory. Peripheral blood specimens were processed by dextran sed