Rapid detection of cytomegalovirus in clinical specimens by using biotinylated DNA probes and analysis of cross-reactivity with herpes simplex virus

Abstract

A method for rapid identification of human cytomegalovirus (HCMV) was developed with biotinylated DNA probes. BamHI restriction fragments from HCMV strain AD169 were selected and tested for their ability to detect virus in patient urine samples. All probes detected 30 pg of HCMV AD169 DNA. The BamHI B fragment detected 15 of 29 cell-culture-positive samples (sensitivity, 52%). There were four samples which were probe positive and cell culture negative (specificity, 87%). The D and H fragments used as combined probes detected 17 of 21 cell-culture-positive samples (sensitivity, 81%). There were five probe-positive and cell-culture-negative samples (specificity, 68.8%). The H fragment, when used alone, detected 11 of 14 culture-positive samples, and 5 samples were culture negative and probe positive. Sensitivity (78.6%) and specificity (76.2%) for the H fragment were similar to those for the combined probes, but the color intensity of the positive reactions detected by the H fragment alone was lower. There was unexpected cross-reactivity with herpes simplex type 1 and 2 controls when the combined D and H probes were used. Specific hybridization was demonstrated between subfragments of the HCMV BamHI D fragment and the herpes simplex virus type 1 EcoRI M fragment.