title: Identification of clinically relevant yeasts by PCR/RFLP.
abstract: For molecular diagnosis of fungal disease using DNA amplification procedures in the routine laboratory, choice of appropriate target structures and rapid and inexpensive identification of amplification products are important prerequisites. Most diagnostic procedures described thus far are characterized by limited applicability, considerable cost for laboratory equipment or low power of discrimination between species. This study aimed at identification of a PCR target appropriate for diagnosis of clinically relevant yeasts and an affordable procedure for characterization of the PCR products to the species level. Here, we describe a PCR-based system using amplification of intergenic spacers ITS1 and ITS2 and restriction length polymorphism of PCR products after sequence-specific enzymatic cleavage. We show the evaluation of the system for clinically relevant Candida species. The simple and inexpensive procedure should be instrumental for rapid identification of medically important yeasts.
authors.affiliation: Institute for Microbiology and Hygiene, Medical Faculty of Humboldt University, Charité, Dorotheenstr. 96, 10117 Berlin, Germany. thomas.adam@charite.de
authors.forename: Anja
authors.initials: A
authors.lastname: Trost
authors.forename: Barbara
authors.initials: B
authors.lastname: Graf
authors.forename: Jan
authors.initials: J
authors.lastname: Eucker
authors.forename: Orhan
authors.initials: O
authors.lastname: Sezer
authors.forename: Kurt
authors.initials: K
authors.lastname: Possinger
authors.forename: Ulf B
authors.initials: UB
authors.lastname: Göbel
authors.forename: Thomas
authors.initials: T
authors.lastname: Adam
mesh_terms: D002175:Candida / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D002177:Candidiasis / Q000382:microbiology*; D017931:DNA Primers / Q000737:chemistry / Q000235:genetics; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D021901:DNA, Intergenic / Q000737:chemistry / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012340:RNA, Ribosomal, 5.8S / Q000737:chemistry / Q000235:genetics
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D021901:DNA, Intergenic; D012340:RNA, Ribosomal, 5.8S