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

• Main Authors: Yong, Phelim Voon Chen, Ong, Khang Wei, Mohd Sidik, Shiran, Rosli, Rozita, Chong, Pei Pei
• Format: Article
• Language: English
• Published: Elsevier BV 2009
• Online Access: http://psasir.upm.edu.my/id/eprint/12728/1/A%20modified%20in%20situ%20RT-PCR%20method%20for%20localizing%20fungal-specific%20gene%20expression%20in%20Candida-infected%20mice%20renal%20cells.pdf; http://psasir.upm.edu.my/id/eprint/12728/

In situ Reverse Transcriptase PCR (in situ RT-PCR) can amplify mRNA and localize gene expression in cells. However, this method is not feasible in fungi as the thick fungal cell wall constitutes a barrier to this procedure. We developed a two step in situ RT-PCR procedure which enabled the detection and localization of Candida tropicalis mRNA expression in formalin-fixed, paraffin-embedded (FFPE) mouse kidney sections. This in situ hybridization study revealed the first direct evidence for deposition of Candida tropicalis secreted aspartic proteinase 2 (CtSAP2) in the tip of pseudohyphae and its involvement in acute systemic candidiasis. We conclude that in situ RT-PCR can be successfully applied to FFPE tissues and will offer new perspectives in studying gene expression in Candida species.