DNA-FINGERPRINTING OF CANDIDA-RUGOSA VIA REPETITIVE SEQUENCE-BASED PCR

A repetitive sequence-based PCR (rep-PCR) technique was developed to c haracterize the genotypic relatedness among Candida rugosa isolates. T wo repetitive sequences, viz., Care-2 and Com29 from Candida albicans, were used to design primers Ca-21, Ca-22, and Com-21, respectively. W hen used alone or in combination, these primers generated discriminato ry fingerprints by amplifying the adjacent variable regions of the gen ome. 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 isola tes into identical groups, indicating that rep-PCR is a specific and r eproducible technique for molecular characterization of C. rugosa. Mor eover, these primers unequivocally discriminated among other important Candida species such as C. albicans, C. glabrata, C. tropicalis, C. k rusei, C. parapsilosis, C. kefyr, and C. lusitaniae. These data confir m the conservation of repetitive sequences in Candida species. Because of its ease and sensitivity,rep-PCR offers a relatively rapid and dis criminatory method for molecular typing of C. rugosa in outbreaks.