Highly polymorphic microsatellite for identification of Candida albicans strains

Abstract

The polymorphism of a new microsatellite locus (CAI) was investigated in a total of 114 C. albicans strains, including 73 independent clinical isolates, multiple isolates from the same patient, isolates from several episodes of recurrent vulvovaginal infections and two reference strains. PCR genotyping was performed automatically, using a fluorescent labelled primer and, in the 73 independent isolates, 26 alleles and 44 different genotypes were identified, resulting in a discriminatory power of 0.97. CAI revealed to be species specific and with a low mutation rate, since no amplification product was obtained when testing other pathogenic Candida species and no genotype differences were observed when testing over 300 generations. When applying this microsatellite to the identification of strains isolated from recurrent vulvovaginal infections in eight patients, it was found that 13 out of 15 episodes were due to the same strain. When multiple isolates obtained from the same patient and plated simultaneously, were typed for CAI, the same genotype was found in each case, confirming that the infecting population was clonal. Moreover, the same genotype appeared in isolates from the rectus and the vagina, revealing that the former could be a reservoir of potentially virulent strains. This new microsatellite proves to be a valuable tool to differentiate C. albicans strains. Furthermore, when compared to other molecular genotyping techniques, CAI proved to be very simple, highly efficient and reproducible, being suitable for low quantity and very degraded samples and for application in large scale epidemiological studies.