All issues > Volume 36(5); 1993

Original Article

J Korean Pediatr Soc. 1993;36(5):626-633. Published online May 15, 1993.

Detection of Cytomegalovirus DNA in Urine Culture Using Polymerase Chain Reaction

Young Mo YM Sohn¹, Won Young WY Lee²

¹Department of Pediatrics, Yonsei University, College of Medicine, Seoul, Korea
²Department of Microbiology, Yonsei University, College of Medicine, Seoul, Korea

Abstract

Polymerase chain reaction(PCR) amplication was used to detect cytomegalovirus (CMV) in tissue culture from the ruine of newborns and patients who was suspected CMV infection. Synthetic oligonucleotide primer pairs were used to amplify DNA from the magor 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 tissue 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 eletrophoresis 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.

Keywords :Congenital cytomegalovirus, Polymerase chain reaction, Hybridization