Evaluation of possibilities of genotyping of Candida glabrata clinical isolates with RAPD-PCR method and microsatellite analysis
نویسندگان
چکیده
The aim of the study was to compare the discriminatory power of RAPD-PCR method using RSD10 primer and microsatellite (GACA)4 analysis for genotyping clinical isolates of C. glabrata. Isolates were received from patients of two Polish hospitals: Children's Memorial Health Institute in Warsaw (n=17) and Medical University of Gdansk (n=37). Species identification was confirmed by two phenotypic methods: growth on chromagar plates (Biocorp) and Candida API ID 32C test (bioMérieux), as well as by PCR reaction. For both tested methods for genotyping, the size of amplified DNA fragments varied mainly between 200 and about 3000 bp. The sets of 9 and 8 different products of amplification were obtained in the case of microsatellite and RAPD-PCR method, respectively. The amplicon arrays were analyzed and dendrograms were constructed using a matrix generated by UPGMA (Unweighted Pair Group Method with Arithmetic Means) in MVSP program (Multi Variate Statistical Package) version 3.22. As a result 20 different genotypes were distinguished in the case of microsatellite analysis and the collection of strains was divided into 26 genotypes in the case of RAPD-PCR analysis with RSD10 primer. Additionally, an evident correlation between genotype classification and geographical origin of the
منابع مشابه
Microsatellite marker analysis as a typing system for Candida glabrata.
Candida glabrata is one of the most important causes of nosocomial fungal infection. We investigated, using a multiplex PCR, three polymorphic microsatellite markers, RPM2, MTI, and ERG3, in order to obtain a rapid genotyping method for C. glabrata. One set of primers was designed for each locus, and one primer of each set was dye labeled to read PCR signals using an automatic sequencer. Eight ...
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