title: A modified in situ RT-PCR method for localizing fungal-specific gene expression in Candida-infected mice renal cells.
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.
authors.affiliation: Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, University Putra Malaysia, Serdang, Selangor, Malaysia.
authors.forename: Voon Chen
authors.initials: VC
authors.lastname: Yong
authors.forename: Khang Wei
authors.initials: KW
authors.lastname: Ong
authors.forename: Shiran Mohd
authors.initials: SM
authors.lastname: Sidik
authors.forename: Rozita
authors.initials: R
authors.lastname: Rosli
authors.forename: Pei Pei
authors.initials: PP
authors.lastname: Chong
mesh_terms: D000818:Animals; D057055:Aspartic Acid Proteases / Q000235:genetics; D041022:Candida tropicalis / Q000235:genetics* / Q000472:pathogenicity; D002177:Candidiasis / Q000382:microbiology*; D005260:Female; D015870:Gene Expression; D005800:Genes, Fungal; D017403:In Situ Hybridization / Q000379:methods*; D007668:Kidney / Q000382:microbiology; D007674:Kidney Diseases / Q000382:microbiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010336:Pathology / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D057055:Aspartic Acid Proteases