Related Experiment Video

Updated: Jun 20, 2026

Visualization of Candida albicans in the Murine Gastrointestinal Tract Using Fluorescent In Situ Hybridization
10:08

Visualization of Candida albicans in the Murine Gastrointestinal Tract Using Fluorescent In Situ Hybridization

Published on: November 5, 2019

A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells

Voon Chen Yong1, Khang Wei Ong, Shiran Mohd Sidik

  • 1Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University Putra Malaysia, Serdang, Selangor, Malaysia.

Journal of Microbiological Methods
|September 10, 2009
PubMed

Insights

We developed a novel in situ RT-PCR method to detect gene expression in fungi. This technique successfully localized Candida tropicalis mRNA and CtSAP2 protein in mouse kidney tissues, advancing fungal research.

Area of Science:

  • Mycology
  • Molecular Biology
  • Infectious Diseases

Background:

  • In situ Reverse Transcriptase PCR (in situ RT-PCR) is crucial for localizing gene expression within cells.
  • The thick fungal cell wall presents a significant barrier, hindering the application of standard in situ RT-PCR in fungi.
  • Developing methods to overcome this barrier is essential for studying fungal gene expression.

Purpose of the Study:

  • To develop and validate a modified in situ RT-PCR technique applicable to fungi.
  • To detect and localize messenger RNA (mRNA) expression of Candida tropicalis in infected tissues.
  • To investigate the role of Candida tropicalis secreted aspartic proteinase 2 (CtSAP2) in systemic candidiasis.

Main Methods:

  • A two-step in situ RT-PCR procedure was developed to enable mRNA detection in fungal cells.
  • Formalin-fixed, paraffin-embedded (FFPE) mouse kidney sections infected with Candida tropicalis were used.
  • In situ hybridization was employed to visualize the localization of CtSAP2 mRNA and protein.

Main Results:

  • The developed two-step in situ RT-PCR successfully detected and localized Candida tropicalis mRNA in FFPE mouse kidney sections.
  • Direct evidence was obtained for the deposition of CtSAP2 at the tip of pseudohyphae.
  • This finding suggests CtSAP2's involvement in the pathogenesis of acute systemic candidiasis.

Conclusions:

  • The modified in situ RT-PCR method is effective for studying gene expression in FFPE fungal tissues.
  • This technique provides new avenues for investigating gene expression in Candida species.
  • The study highlights the role of CtSAP2 in Candida tropicalis infections.

More Related Videos

Gene Expression Profiling of Infecting Microbes Using a Digital Bar-coding Platform09:13

Gene Expression Profiling of Infecting Microbes Using a Digital Bar-coding Platform

Published on: January 13, 2016

Related Concept Videos

Real Time RT-PCR02:57

Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...