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KMID : 0603520010060020085
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Journal of Korean Association of Cancer Prevention
2001 Volume.6 No. 2 p.85 ~ p.91
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Detection and Sequence Analysis of DNA Flanking Integrated Hepatitis B Virus in Peripheral Blood Mononuclear Cells by Three-Step PCR
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Kim Woo-Jean
Lee Sae-Won
Park Neung-Hwa
Kim Kyu-Won
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Abstract
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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.
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KEYWORD
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Hepatitis B virus, Peripheral blood mononuclear cells
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