Identification of clinically relevant yeasts by PCR/RFLP.
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.
Trost
Anja
A
Institute for Microbiology and Hygiene, Medical Faculty of Humboldt University, Charité, Dorotheenstr. 96, 10117 Berlin, Germany. thomas.adam@charite.de
Graf
Barbara
B
Eucker
Jan
J
Sezer
Orhan
O
Possinger
Kurt
K
Göbel
Ulf B
UB
Adam
Thomas
T
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
D017931:DNA Primers; D004271:DNA, Fungal; D021901:DNA, Intergenic; D012340:RNA, Ribosomal, 5.8S