Yonago Acta medica 1996;39:171–176
Improvements in the Detection of Epstein-Barr Virus DNA on Paraffin-Embedded Gastric Carcinoma Tissues: Treatment of Extracted Cellular DNA with a Restriction Enzyme Prior to Polymerase Chain Reaction
Noriko Takasaka, Yukio Satoh, Yoshiko Hoshikawa, Mitsuhiko Osaki*, Hisao Ito*, Jen-Yang Chen† and Takeshi Sairenji
Department of Biosignaling and *First Department of Pathology, Faculty of Medicine, Tottori University, Yonago 683 and †Department of Bacteriology and Graduate Institute of Microbiology, College of Medicine, National Taiwan University, Taipei, Taiwan
An association between Epstein-Barr virus (EBV) and a certain gastric carcinoma has been suggested by the presence of EBV-DNA in the carcinoma cells. We tried to detect EBV-DNA in paraffin-embedded tissues of gastric carcinoma by polymerase chain reaction (PCR) followed by Southern analysis. The EBV-DNA sequence was not amplified sufficiently with a BamH I-W primer pair to the extracted DNA itself from the tissues which were detected for the expression of EBV-encoded small RNA (EBER), while the sequence was amplified to the DNA from Raji cells as a positive control. However, when the extracted DNA was digested with a restriction enzyme, BamH I or EcoR I and then subjected to PCR, the BamH I-W region was amplified in the DNA from EBER-positive but not -negative tissues. In the extracted DNA from an EBV genome-carrying cell line derived from a gastric carcinoma, the amplification with PCR was increased slightly by the digestion with BamH I. Thus, the pretreatment for template DNA increased the sensitivity of PCR for the detection of EBV-DNA from paraffin-embedded tissues.
Key words: Epstein-Barr virus; gastric carcinoma; PCR; restriction enzyme
