EVALUATION AND COMPARISON OF PCR AND HYBRIDIZATION METHODS FOR RAPID DETECTION OF CYTOMEGALOVIRUS IN CLINICAL-SAMPLES

Rapid diagnosis of cytomegalovirus (CMV) infection may be obtained by molecular techniques, such as the polymerase chain reaction (PCR) and hybridization assays. The optimal technique to detect CMV in clinical samples was assessed. Two different PCR assays were used, targeting ei ther the major immediate early 1 (MIE 1) or the HXLF 4 gene. The PCR p roducts were detected by gel electrophoresis, dot blotting and an easy to use; rapid, solid phase hybridization assay, DNA enzyme immunoassa y (DEIA). Standard tissue culture was also used. Cerebrospinal fluids (18), liver biopsies (9) from hepatic transplant recipients, amniotic fluids (7) from mothers with suspected peripartum infection, and sampl es (6) of miscellaneous origin (brain and fundus biopsy, pericardial a nd pleural fluid) were tested. Among the 40 samples, CMV was detected in 19 cases. Three were positive by both molecular techniques and tiss ue culture, 14 by molecular methods and 2 by culture. 16/19 or 9/19 CM V-positive samples were detected by PCR amplification of the HXLF 4 or MIE 1 gene, respectively and 14/16 HXLF 4-positive samples were detec ted using either dot-blot or DEIA, compared to 9/16 using gel electrop horesis. Thus, the most sensitive assays for the detection of CMV in c linical samples using the methods compared in the current study were P CR amplification of the HXLF 4 gene followed by dot-blot or DEIA hybri dization.