Comparison between the Effectiveness of Polymerase Chain Reaction and In Situ Hybridization in Detecting the Presence of Pathogenic Fungi by Using the Preserved DNA in Formalin-Fixed and Paraffin-Embedded Tissues
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概要
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In situ hybridization (ISH) has been recognized as an important technique for identifying the causative fungi in the foci of infection observed in histopathological specimens which was processed from formalin-fixed and paraffin-embedded (FFPE) tissues. However, few basic studies have conducted an evaluation of the DNA preservation for use in ISH in comparison to polymerase chain reaction (PCR). The latter is a DNA amplification-based modality. In the present study, we analyzed 65 FFPE lung tissue specimens collected from autopsy cases for comparing the usefulness of ISH and PCR analysis. As a result, the positive identification rates for PCR were strikingly low; a majority of these results can be assumed to be false negative because the presence of fungi had been confirmed by histopathological analysis. In contrast, panfungal ISH targeting of the 28S rRNA showed a higher sensitivity than the 230-bp panfungal PCR primers did (80.0% versus 4.6%, respectively). Furthermore, over 60% of the samples we examined showed a favorable intensity of the ISH signal. Therefore, in conventional postmortem FFPE tissues, the state of DNA preservation may be more favorable for ISH than PCR analysis.
著者
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Ide Tadashi
Department Of Surgical Pathology Toho University School Of Medicine
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Yamagata Murayama
Laboratory Of Molecular Epidemiology For Infectious Agents Graduate School Of Infection Control Scie
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Nakayama Haruo
Department Of Material Science Faculty Of Engineering Yokohama National University
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Shibuya Kazutoshi
Department Of Clinical Pathology School Of Medicine Toho University
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Wakayama Megumi
Department Of Pathology Ohashi Hospital Toho University School Of Medicine
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Shinozaki Minoru
Department Of Surgical Pathology Toho University School Of Medicine
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Okubo Yoichiro
Department Of Surgical Pathology Toho University Omori Medical Center
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Zhang Zean
Center for Drug Safety Evaluation and Research, Shanghai University of T.C.M., 1200 Cailun Road, Shanghai 201203, China
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Zhi Yuzhu
Department of Surgical Pathology, Toho University School of Medicine
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Sasai Daisuke
Department of Surgical Pathology, Toho University School of Medicine
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Zhang Zean
Center for Drug Safety Evaluation
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Sasai Daisuke
Department of Surgical Pathology, Toho University Omori Medical Center
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YAMAGATA MURAYAMA
Laboratory of Molecular Cell Biology, School of Pharmacy, Nihon University
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Nakayama Haruo
Department of Applied Chemistry, Faculty of Engineering, Yokohama National University
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