We found a match
Your institution may have rights to this item. Sign in to continue.
- Title
Application of In Situ Hybridization to Tissue Sections for Identification of Molds Causing Invasive Fungal Infection.
- Authors
Shinozaki, Minoru; Okubo, Yoichiro; Nakayama, Haruo; Mitsuda, Aki; Ide, Tadashi; Murayama, Somay Yamagata; Shibuya, Kazutoshi
- Abstract
The present article describes our studies to know the usefulness of in situ hybridization (ISH) to identify various kinds of mold observed in tissue sections and/or cytological preparations from the lesions of patients with invasive fungal infection. To establish the precise procedure for ISH in formalin-fixed and paraffin- embedded sections, various pretreatments were attempted. The condition finally chosen is written here providing a favorable outcome regarding to both intensity and specificity of signals on outline of molds observed in the tissue sections when specimens were treated with both heat and proteinase K and, solutions were adjusted to higher pH value. Therefore, usefulness of promising probes, two each DNA and peptide nucleic acid (PNA) were verified with a favorable pretreatment condition, using lungs of mice experimentally infected and/or those obtained from autopsies with invasive mold infection. As the result, DNA probes targeting alkaline proteinase (ALP) gene and retrotransposon Afut-1 gene of Aspergillus furnigatus showed specific signal intensity for the Aspergillus species and A. fumigatus, respectively. PNA probes for Candida albicans and the Fusarium species also showed satisfactory specificity. We wish to emphasize that ISH can be a valuable tool to identify medically important molds in formalin-fixed and paraffin-embedded tissue sections or cytological preparations.
- Subjects
IN situ hybridization; NUCLEIC acid hybridization; MOLDS (Fungi); MYCOSES; PROTEINASES; NUCLEIC acids; LABORATORY animals
- Publication
Japanese Journal of Medical Mycology, 2009, Vol 50, Issue 2, p75
- ISSN
0916-4804
- Publication type
Article
- DOI
10.3314/jjmm.50.075