Cytomeglovirus is the most common cause of congenital and perinatal infections throughout the world. The early diagnosis of CMV infection provides the opportunity for identification of asymptomatic infants for follow-up and intervention. The standard procedure for diagnosis is isolation of the virus from urine by tissue culture. However, culture is not rapid and widely available. Polymerase chain reaction (PCR) offers several advantages over culture but requires confirmation of amplicon identity. In order to evaluate a simplified and more rapid PCR method we tested a multiplex PCR for diagnosis of congenital and perinatal CMV infection comparing it to fibroblast CMV culture. Fifty urine specimens of newborns and infants were tested. The PCR was carried out using three primer pairs simultaneously (MIE-producing a 435 base pair (bp) amplicon, part of the gene codifying the major immediate-early virus proteins; gB - producing a 296 bp amplicon of glycoprotein B gene; gH - producing a 215 bp amplicon of glycoprotein H gene). The multiplex PCR was considered positive if products of at least 2 primer pairs were amplified. Forty one of the 44 positive samples by CMV culture isolation were positive by multiplex PCR including 38 samples with 3 amplicons and 3 samples with 2 amplicons. The remaining 3 of these 44 urine samples were negative by multiplex PCR. No amplified product was obtained by multiplex PCR with the 6 negative urine samples. The multiplex PCR compared to virus isolation (gold standard) showed 93.3% sensitivity and 100% specificity. The PCR multiplex dispenses further procedures for confirmation of amplicon identify because each amplicon provides an internal control for the other amplified fragments.

We conclude that this CMV multiplex assay in urine is rapid, simple and has a suitable sensitivity for the diagnosis of congenital and perinatal infections.