PCR을 이용한 인간 세포거대바이러스(HCMV) DNA 중합효소 유전자의 검색

Abstract

Polymerase chain reaction(PCF) amplification was used to detect human 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 control. Amplified products were 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 10²³TCID₅₀/0.1㎖ of cell culture muxture was detected by direct agarose gel electrophoresis;South-ern blot hybridization was four-fold more sensitive(10¹·⁷TCID₅₀/0.1㎖) than gel analysis. The specificity of the PCR was evaluated using other members of the herpes family of viruses and various DANs. No amplification was noted by direct gel analysis or by dot blot hybridization and only HCMV containing specimen was positive. In conclusion, these results showed that the PCR may be a sensitive and fast tool for the diagno-sis of HCMV infections. Key Words : Human Cytomegalovirus(HCMV), DNA polymerase gene, PCR, Southern blot.