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KMID : 0357319920270020181
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Journal of the Korean Society for Microbiology
1992 Volume.27 No. 2 p.181 ~ p.188
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Decection of Human Cytomegalovirus DNA Polymerase Gene by Polymerase Chain Reaction
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김현철
박성배/조원현/백원기/서민호
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Abstract
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Polymerase chain reaction (PCR) amplificatin was used to detect hyman cytomegalovirus (HCMV) in MRC-5 cell culture. Oligonucleotide pairs for DNA polymerase gene of HCMV were used as primers and HCMV AD169 strain was used as a cntrol. Amplified.
Amplified products detected by gel electrophoresis and by Southern blot hybridization with digoxigenin-labeled HCMV DNA polymerase gene probe. In estimating the sensitivity of PCR, a 5㎍ aliquot of an infectivity titer of 10E2.3 TCID 50/0.1ml of
cell
culture muxture was detected by direct agarose gel electrophoresis ; Southern blot hybridization was four-fold more sensitive(10E1.7 TCID50/0.1ml) than gel analysis.
The specificity of the PCR was evaluated using other members of the herpes family of viruses and various DNAs. No amplification was noted by direct gel analysis or by dot blot hybridizationand only HCMV containing specimen was positive.
In conclusion, these results showed that the PCR may be a sensitive and fast tool for the diagnosis of HCMV infections.
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