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Robust Detection of Gene Amplification in Formalin-Fixed Paraffin-Embedded Samples by Fluorescence In Situ Hybridization
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Identification of Fusarium species in formalin-fixed and paraffin-embedded sections by in situ hybridization using

Minoru Shinozaki1, Yoichiro Okubo, Daisuke Sasai

  • 1Department of Surgical Pathology, Toho University School of Medicine, 6-11-1 Omori-Nishi, Ota-Ku, Tokyo 143-8541, Japan.

Journal of Clinical Microbiology
|November 26, 2010
PubMed

Insights

Histological differentiation of Fusarium from similar fungi is challenging. In situ hybridization (ISH) with peptide nucleic acid (PNA) probes accurately identifies Fusarium species in tissue sections, aiding pathological diagnosis.

Area of Science:

  • Medical Mycology
  • Pathology
  • Molecular Diagnostics

Background:

  • Fusarium species are emerging opportunistic human pathogens.
  • Histological differentiation of Fusarium from Aspergillus and Scedosporium is difficult due to similar clinical and morphological features.
  • There is a need for rapid and reliable diagnostic methods for Fusarium infections.

Purpose of the Study:

  • To evaluate and establish in situ hybridization (ISH) using peptide nucleic acid (PNA) probes for Fusarium identification in tissue sections.
  • To target the 28S ribosomal RNA (rRNA) gene for specific detection of Fusarium species.

Main Methods:

  • Validation of ISH with PNA probes using formalin-fixed, paraffin-embedded pulmonary tissues from experimentally infected mice.
  • Testing on cell blocks from fungal cultures of 30 strains, including seven fungal species.
  • Assessment of signal intensity and cross-reactivity with other fungal species.

Main Results:

  • Strong positive signals were observed in lung tissues infected with Fusarium solani.
  • The PNA probe showed strong reactivity with Fusarium solani and Fusarium oxysporum in cell block sections.
  • Low-intensity cross-reactivity was noted with Pseudallescheria boydii, but most hyphae were non-reactive.

Conclusions:

  • In situ hybridization (ISH) using PNA probes targeting 28S rRNA is accurate for identifying Fusarium species.
  • This method is a valuable tool for differentiating Fusarium from morphologically similar fungi in pathological specimens.
  • The technique aids in accurate diagnosis of Fusarium infections in formalin-fixed, paraffin-embedded tissues.

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