Detection of Candida albicans mRNA from formalin-fixed, paraffin-embedded mouse tissues by nested reverse transcription-PCR.
Histopathology archives represent a vast source of infectious disease specimens that can be used to elucidate important disease processes. In this report, we describe a method to detect Candida albicans gene expression from infected, formalin-fixed, paraffin-embedded mouse tissue. By use of glass beads to break open the fungal cells and proteinase K treatment, RNA was extracted routinely from tissue sections that had been fixed for up to 72 h. Upon reverse transcription of the RNA and nested PCR, the procedure detected C. albicans "housekeeping" and putative virulence genes.
Schofield
David A
DA
Department of Microbiology and Immunology, Medical University of South Carolina, Charleston, South Carolina 29403, USA. schofida@musc.edu
Westwater
Caroline
C
Paulling
Emily E
EE
Nicholas
Peter J
PJ
Balish
Edward
E
D000818:Animals; D002176:Candida albicans / Q000235:genetics / Q000302:isolation & purification*; D005557:Formaldehyde / Q000737:chemistry; D006652:Histological Techniques; D051379:Mice; D016612:Paraffin Embedding; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
D012333:RNA, Messenger; D005557:Formaldehyde