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In-situ detection of Aspergillus fumigatus

RYO HANAZAWA^*, SOMAY YAMAGATA MURAYAMA^* and HIDEYO YAMAGUCHI^*,

*Department of Microbiology and Immunology, Teikyo University School of Medicine, 2-11-1, Kaga, Itabashi-Ku, Tokyo 173-8605 and Teikyo University Institute of Medical Mycology, 359, Otsuka, Hachioji, Tokyo 192-0395, Japan

Corresponding author: Dr R. Hanazawa (e-mail: rhanazaw{at}med.teikyo-u.ac.jp).

Received 23 Feb. 1999; revised version accepted 8 Aug. 1999; accepted 16 Aug. 1999.

Abstract

An in-situ hybridisation (ISH) technique to detect Aspergillus fumigatus in infected tissues was developed in which 568-bp, 333-bp and 154-bp PCR products of the alkaline proteinase gene were employed. Dot-blot hybridisation with the 568-bp probe on a membrane containing genomic DNA from several different fungi including A. flavus, A. niger, Penicillium spp., Mucor racemosus or Pseudallescheria boydii gave negative results. ISH was done on formalin-fixed, paraffin-embedded pulmonary tissues from rats infected with A. fumigatus and renal tissues from mice infected with A. fumigatus, A. flavus or A. niger. The 568-bp probe reacted strongly in ISH with both A. fumigatus and A. flavus, and weakly with A. niger. The 333-bp probe also reacted in ISH with A. fumigatus and A. flavus, although the intensity was weaker. However, in ISH with the 154-bp probe, there was no positive signal with any Aspergillus spp. These results demonstrate that A. fumigatus and A. flavus can be specifically detected in infected tissues by ISH with the 568-bp probe. This technique could be applicable to clinical specimens for molecular diagnosis of aspergillus infections.

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