Document Detail


Applications of differential-display reverse transcription-PCR to molecular pathogenesis and medical mycology.
MedLine Citation:
PMID:  10885984     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The host-fungus interaction is characterized by changes in gene expression in both host and pathogen. Differential-display reverse transcription PCR (DDRT-PCR) is a PCR-based method that allows extensive analysis of gene expression among several cell populations. Several limitations and drawbacks to this procedure have now been addressed, including the large number of false-positive results and the difficulty in confirming differential expression. Modifications that simplify the reaction time, allow the use of minute quantities of RNA, or address unusual species- or gene-specific sequences have been reported. DDRT-PCR has been used to address biological questions in mammalian systems, including cell differentiation, cell activation, cell stress, and identification of drug targets. In microbial pathogenesis and plant pathogenesis, DDRT-PCR has allowed the identification of virulence factors, genes involved in cell death, and signaling genes. In Candida albicans, DDRT-PCR studies identified TIF-2, which may play a role in the upregulation of phospholipases, and the stress-related genes, CIP1 and CIP2. In Histoplasma capsulatum and C. albicans, genes involved in the host-pathogen interaction, including a member of the 100-kDa family in Histoplasma and an ALS and 14-3-3 gene in Candida, were potentially identified by DDRT-PCR. Although very few reports have been published in medical mycology, studies in mammalian, nonfungal microbial, and plant pathogen systems are easily applied to basic questions in fungal pathogenesis and antifungal therapeutics.
Authors:
J Sturtevant
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Publication Detail:
Type:  Journal Article; Review    
Journal Detail:
Title:  Clinical microbiology reviews     Volume:  13     ISSN:  0893-8512     ISO Abbreviation:  Clin. Microbiol. Rev.     Publication Date:  2000 Jul 
Date Detail:
Created Date:  2000-08-03     Completed Date:  2000-08-03     Revised Date:  2013-04-17    
Medline Journal Info:
Nlm Unique ID:  8807282     Medline TA:  Clin Microbiol Rev     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  408-27     Citation Subset:  IM    
Affiliation:
Department of Microbiology, Georgetown University Medical School, Washington, DC 20007, USA. sturtevj@gusun.georgetown.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Cycle
Cell Differentiation
Cloning, Molecular
Gene Expression
Humans
Mycology*
Reverse Transcriptase Polymerase Chain Reaction / methods*
Virulence
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