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Detection of Cytomegalovirus DNA in Urine Culture Using Polymerase Chain Reaction

Journal of the Korean Pediatric Society 1993;36(5):626-633.
Published online May 15, 1993.
Detection of Cytomegalovirus DNA in Urine Culture Using Polymerase Chain Reaction
Young Mo Sohn1, Won Young Lee2
1Department of Pediatrics, Yonsei University, College of Medicine, Seoul, Korea
2Department of Microbiology, Yonsei University, College of Medicine, Seoul, Korea
중합효소연쇄반응을 이용한 소변배양에서 거대세포바이러스 DNA 검출
손영모1, 이원영2
1연세대학교 의과대학 소아과학교실
2연세대학교 의과대학 미생물학교실
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.
Key Words: Congenital cytomegalovirus, Polymerase chain reaction, Hybridization


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