인간 세포거대바이러스의 배양상 및 PCR에 의한 바이러스 DNA 검색
- 인간 세포거대바이러스의 배양상 및 PCR에 의한 바이러스 DNA 검색
- Other Titles
- Cultural Characteristics of Human Cytomegalovirus and Detection of Viral DNA by Polymerase
- 박건욱; 백성덕; 백원기; 서성일; 박종욱; 서민호; 최병길; Keon-Wook Park; Seong-Duck Paik; Won-Ki Baek; Seong-Il Suh; Jong-Wook Park; Min-Ho Suh; Byung-Kil Choe
- Keimyung Author(s)
- 백원기; 서성일; 서민호; 박종욱; 최병길
- Dept. of Microbiology (미생물학); Dept. of Immunology (면역학)
- Human Cytomegalovirus (HCMV); Culture; Major Immediate Early gene(MIE); PCR
- Issue Date
- School of Medicine
- 대한미생물학회 The Korean Society For Microblology, Vol.29(3) : 275-285, 1994
- In order to understand the cultural characteristics of human cytomegalovirus (HCMV) and to make direct detection of viral DNA during cell culture, the viruses were cultured in MRC-5 fibroblast cell line and culture supernatants were tested by the Polymerase Chain Reaction (PCR).
Charactistic cytopathic effects (CPE) of HCMV infection were detected after 10 days of virus inoculation. Characteristic intranuclear inclusion bodies and large, round or oval shaped cells were clearly seen after 12 days of virus inoculation.
In PCR, oligonucleotide pairs for major immediate early (MIE) gene of HCMV were used as primers. Amplified products were detected by gel electrophoresis and by Southern blot hybridization with digoxigenin-labeled HCMV MIE gene probe. In estimating the sensitivity of PCR, the PCR product starting with l/i\ aliquot of an infectivity titer of 102 5 TCIDso/0.2ml of cell culture mixture was detedcted by direct agarose gel electrophoresis and Southern blot hybridization. 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 Southern blot hybridization and only HCMV containing specimen was positive. In the sensitivities and the specificities, there were no differences between the PCR with OPC(oligonucleotide purifica¬tion column) purified primers and the PCR with Speed Vac dried primers (no other purification steps were used).
Culture supernatants were tested for the detection of MIE DNA by PCR. After 4 days of culture, the MIE DNA was detected in culture supernatants by PCR.
In conclusion, I think we can make more rapid and precise diagnosis of HCMV infection by combining virus culture and PCR of culture supernatants.
- Appears in Collections:
- 1. 연구논문 > 1. School of Medicine (의과대학) > Dept. of Microbiology (미생물학)
1. 연구논문 > 1. School of Medicine (의과대학) > Dept. of Immunology (면역학)
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