長期ホルマリン固定肺組織からのDNA抽出 : PCR法によるPneumocystis carinii DNAの検出(自然科学系)

概要

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Pneumonia caused by Pneumocystis carinii (Pc) has been diagnosed by direct detection of Pc in sputum and bronchoalveolar lavage (BAL) collected from patients. In recent years, a method using polymerase chain reaction (PCR) has been more frequently applied to diagnosis of Pc pneumonia. As material for detection of Pc by the PCR method, sputum, BAL, and formalin-fixed tissue are used. Pc genes, however, cannot be detected by PCR in specimens fixed in formalin for a prolonged time. In this study, we improved a method of extracting 'DNA from lung specimens fixed in formalin for a prolonged time. As target genes for amplification, ribosomal RNA internal transcribed spacers, 5S ribosomal RNA, and mitochondrial ribosomal RNA of Pc were selected. As the control for confirming that template DNAs Were appropriate for PCR amplification, human β-globin gene was amplified. Using this method of DNA extraction, in which digestive and fixative removal were promoted, the Pc genes were detected in lung tissue specimens fixed in formalin. Amplification of the control human β-globin gene was also improved, suggesting that Pc detection in fixed lung tissue specimens in this study was due to an increased amount of template DNA by improved DNA extraction.
大阪府立大学の論文
2000-03-27