Rapid detection of cytomegalovirus infections by a tissue culture-centrifugation-monoclonal antibody-biotin/avidin immunofluorescence technique

Abstract

A technique using centrifugation, early antigen detection by biotin/avidin fluorescence assay (CEADBAFA) for detection of CMV was compared to a similar method without biotin/avidin (CEADIFA) and to tissue culture alone (TC). The technique using CEADBAFA was the most sensitive of the three assays. The number of CMV-specific fluorescent nuclei was 2-15 fold higher after centrifugation of cell culture monolayers than without centrifugation and the maximum number of positive nuclei was observed after 18 h postinoculation. E-13 was found to be the best of three commercial monoclonal antibodies for identifying CMV positive nuclei. A total of 260 urine, respiratory and genital specimens were tested by both CEADBAFA and TC. Of 46 clinical specimens positive for CMV, 70% were positive by both assays, 7% were positive by TC only and 24% were positive by CEADBAFA only. Two of three specimens positive by TC only could not be evaluated by CEADBAFA because the specimens were toxic to the monolayers on the cover slips. Nearly all of the CEADBAFA positive but TC negative specimens were from patients with a history of CMV infection demonstrated either by TC or serology in other specimens. The CEADBAFA technique should be used for the routine diagnosis of CMV infection.