Detection of cytomegalovirus DNA in urine culture using polymerase chain reaction

Sohn, Young Mo; Lee, Won Young

Abstract

Polymerase chain reaction (PCR) amplication was used to detect cytomegalovirus (CMV) in tissue culture from the urine of newborns and patients who was suspected CMV infection, Synthetic oligonucleotide primer pairs were used to amplify DNA from the major immediate-early and the phosphoprotein 150 genes of CMV AD 169. Amplified products were detected by gel electrophoresis and by dot-blot hybridization with oligonucleotide probes. We found 12 different tissus culture isolates of CMV after the microimmunoassay using monoclonal antibody to immediate-early antigen. All 12 isolates were positive after PCR amplification. But there was no positive reaction when the same primers and probes were used to amplify herpes simplex virus and human genomic DNA. Twelve urine samples were positive when tested with one or both primer pairs and probes. When compaired tissue culture, detection gel electrophoresis provide a sensitivity of 91% (11/12), dot-blot analysis raised the sensitivity to 100% (12/12). A specificity of both primer was 100%(0/12). We conclude that PCR amplification may be a valuable tool for diagnosing congenital CMV infection.