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

Citation

Yong, Phelim Voon Chen and Ong, Khang Wei and Mohd Sidik, Shiran and Rosli, Rozita and Chong, Pei Pei (2009) A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells. Journal of Microbiological Methods, 79 (2). pp. 242-245. ISSN 0167-7012; ESSN: 1872-8359

Abstract

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.

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Additional Metadata

Item Type:	Article
Divisions:	Faculty of Medicine and Health Science
DOI Number:	https://doi.org/10.1016/j.mimet.2009.08.019
Publisher:	Elsevier BV
Keywords:	Candida tropicalis; In situ RT-PCR; Secreted aspartyl proteinase; Biotinylated dUTP; Gene expression
Depositing User:	Nurul Ainie Mokhtar
Date Deposited:	29 May 2015 05:51
Last Modified:	03 Nov 2015 01:45
Altmetrics:	http://www.altmetric.com/details.php?domain=psasir.upm.edu.my&doi=10.1016/j.mimet.2009.08.019
URI:	http://psasir.upm.edu.my/id/eprint/12728
Statistic Details:	View Download Statistic

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