In-situ detection of Aspergillus fumigatus.
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.
Hanazawa
Ryo
R
*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.
Murayama
Somay Yamagata
SY
*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.
Yamaguchi
Hideyo
H
*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.
D000818:Animals; D001228:Aspergillosis / Q000382:microbiology*; D001232:Aspergillus fumigatus / Q000201:enzymology / Q000235:genetics / Q000302:isolation & purification*; D015342:DNA Probes / Q000592:standards; D004271:DNA, Fungal / Q000032:analysis*; D010450:Endopeptidases / Q000235:genetics*; D005260:Female; D017403:In Situ Hybridization; D007668:Kidney / Q000382:microbiology; D008168:Lung / Q000382:microbiology; D051379:Mice; D008813:Mice, Inbred ICR; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D013045:Species Specificity; D013047:Specific Pathogen-Free Organisms
D015342:DNA Probes; D004271:DNA, Fungal; D010450:Endopeptidases