말초혈액 단핵구에서 3단계 PCR에 의한 B형 간염 바이러스 삽입 부위 탐색
Detection and Sequence Analysis of DNA Flanking Integrated Hepatitis B Virus in Peripheral Blood Mononuclear Cells by Three-Step PCR
김우진, 이세원, 박능화, 김규원,
소속 상세정보
김우진 ( Kim Woo-Jean )
부산대학교 분자생물학과
이세원 ( Lee Sae-Won )
부산대학교 분자생물학과
박능화 ( Park Neung-Hwa )
울산의대 울산대학교병원 내과학교실
김규원 ( Kim Kyu-Won )
서울대학교 약학대학
KMID : 0603520010060020085
Abstract
Hepatitis B virus (HBV)-DNA insertion into the human genome was highly related with acute-or chronic- human disease. To diagnose the HBV-DNA, several methods have been introduced to investigate the viral integration site. However, these methods were so complex and expensive. Thereby we modified the rapid amplification of cDNA ends (RACE) into three-step PCR technique that can simply figure out the flanking region or viral integration sites. By using three-step PCR, we detected HBV-cellural DNA junctions in all 3 among 13 patients. The HBV flanking region was cytosine-rich or guanine-rich. The pyrimidine or cytosine-rich region was known as a suitable environment for acting of topoisomerase Ⅰ that is related with HBV integration. The fact HBV integration into peripheral blood mononuclear cells (PBMC) showed on possibility that immune system will be damaged directly by HBV infection in PBMC. From these results, we suggest that three-step PCR is a useful method to figure out and characterize the viral integration sites including HBV integration sites.
키워드
Hepatitis B virus; Peripheral blood mononuclear cells
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