DNA fingerprinting of Candida rugosa via repetitive sequence-based PCR.
A repetitive sequence-based PCR (rep-PCR) technique was developed to characterize the genotypic relatedness among Candida rugosa isolates. Two repetitive sequences, viz., Care-2 and Com29 from Candida albicans, were used to design primers Ca-21, Ca-22, and Com-21, respectively. When used alone or in combination, these primers generated discriminatory fingerprints by amplifying the adjacent variable regions of the genome. Twenty-three isolates from burn patients, eight from other human sources, and four C. rugosa isolates pathogenic in animals were placed into nine fingerprinting groups. Different primers placed these isolates into identical groups, indicating that rep-PCR is a specific and reproducible technique for molecular characterization of C. rugosa. Moreover, these primers unequivocally discriminated among other important Candida species such as C. albicans, C. glabrata, C. tropicalis, C. krusei, C. parapsilosis, C. kefyr, and C. lusitaniae. These data confirm the conservation of repetitive sequences in Candida species. Because of its ease and sensitivity, rep-PCR offers a relatively rapid and discriminatory method for molecular typing of C. rugosa in outbreaks.
Redkar
R J
RJ
Molecular Biology Institute, University of Scranton, Pennsylvania 18510, USA.
Dubé
M P
MP
McCleskey
F K
FK
Rinaldi
M G
MG
Del Vecchio
V G
VG
D000818:Animals; D001483:Base Sequence; D002175:Candida / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D002176:Candida albicans / Q000235:genetics; D002177:Candidiasis / Q000453:epidemiology / Q000382:microbiology; D016172:DNA Fingerprinting / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004196:Disease Outbreaks; D005069:Evaluation Studies as Topic; D005838:Genotype; D006801:Humans; D017720:Molecular Epidemiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000706:statistics & numerical data; D012091:Repetitive Sequences, Nucleic Acid; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D004271:DNA, Fungal