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Supplemental utility of nested PCR for the pathological diagnosis of disseminated trichosporonosis

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Abstract

Disseminated trichosporonosis is known to be a severe opportunistic mycosis and has a high mortality rate. In autopsy cases, it is often difficult to diagnose as trichosporonosis because the causative Trichosporon species are pathologically similar to other fungi, especially the Candida species. Immunohistochemical analysis is essential for the differential diagnosis, but an antibody to Trichosporon is not available commercially. In the present study, we investigated the supplemental utility of nested polymerase chain reaction (PCR) for the pathological diagnosis of trichosporonosis from formalin-fixed and paraffin-embedded tissues. Total DNA was purified from 30 major organs in three autopsy cases, and Trichosporon DNA was specifically amplified by nested PCR using three sets of primers. Of 22 organs in which Grocott’s stain was positive for fungal infection, 170- and 259-bp PCR products were detected in 20 (91%) and 12 (55%) organs, respectively. In short-term fixation (about 1 day), these bands were highly detected in ten (100%) and nine (90%) organs, whereas the detection efficiency tended to decrease after long-term fixation and decalcification. No PCR product of 412 bp was detected in any organs. These findings suggest that nested PCR from short-term-fixed tissues is useful for supportive pathological diagnosis of disseminated trichosporonosis.

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Acknowledgment

The authors thank Dr. Masao Hotchi, Professor Emeritus, Shinshu University for excellent immunohistochemistry and useful discussion; Prof. Jun-ichi Kadota and Dr. Issei Tokimatsu, Division of Pathogenesis and Disease Control, Department of Infectious Diseases, Oita University Faculty of Medicine, and Dr. Shiro Naoe, Professor Emeritus, Toho University for their critical suggestion and discussion; Prof. Tsutomu Katsuyama, Laboratory Medicine, Shinshu University Hospital for kindly advise; and Ms. Michiko Yagoshi, Clinical Laboratory, Nihon University Itabashi Hospital, and Ms. Nozomi Watanabe for helpful information on the Trichosporon species.

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Authors and Affiliations

  1. Department of Pathology, Nihon University School of Medicine, 30-1 Oyaguchi-Kamimachi, Itabashi-ku, Tokyo, 173-8610, Japan

    Makoto Sano, Masahiko Sugitani, Toshiyuki Ishige, Taku Homma, Kentaro Kikuchi, Keishin Sunagawa, Yukari Obana & Norimichi Nemoto

  2. Department of Laboratory Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-Kamimachi, Itabashi-ku, Tokyo, 173-8610, Japan

    Yuki Uehara & Kazunari Kumasaka

  3. Division of Hematology and Rheumatology, Department of Medicine, Nihon University School of Medicine, 30-1 Oyaguchi-Kamimachi, Itabashi-ku, Tokyo, 173-8610, Japan

    Kumi Uenogawa, Sumiko Kobayashi, Yoshihiro Hatta & Jin Takeuchi

  4. Division of Infectious Diseases, Department of Medicine, St. Luke’s International Hospital, 9-1 Akashicho, Chuo-ku, Tokyo, 104-8560, Japan

    Yuki Uehara

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  1. Makoto Sano
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  2. Masahiko Sugitani
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  3. Toshiyuki Ishige
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Correspondence to Makoto Sano.

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Sano, M., Sugitani, M., Ishige, T. et al. Supplemental utility of nested PCR for the pathological diagnosis of disseminated trichosporonosis. Virchows Arch 451, 929–935 (2007). https://doi.org/10.1007/s00428-007-0484-6

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  • DOI: https://doi.org/10.1007/s00428-007-0484-6

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