Abstract
BackgroundWe have previously reported the use of PCR Melting Profile (PCR MP) technique based on using low denaturation temperatures during ligation mediated PCR (LM PCR) for bacterial strain differentiation. The aim of the current study was to evaluate this method for intra-species differentiation of Candida albicans strains.MethodsIn total 123 Candida albicans strains (including 7 reference, 11 clinical unrelated, and 105 isolates from patients of two hospitals in Poland) were examined using three genotyping methods: PCR MP, macrorestriction analysis of the chromosomal DNA by pulsed-field gel electrophoresis (REA-PFGE) and RAPD techniques.ResultsThe genotyping results of the PCR MP were compared with results from REA-PFGE and RAPD techniques giving 27, 26 and 25 unique types, respectively. The results showed that the PCR MP technique has at least the same discriminatory power as REA-PFGE and RAPD.ConclusionData presented here show for the first time the evaluation of PCR MP technique for candidial strains differentiation and we propose that this can be used as a relatively simple and cheap technique for epidemiological studies in short period of time in hospital.
Highlights
We have previously reported the use of PCR Melting Profile (PCR MP) technique based on using low denaturation temperatures during ligation mediated PCR (LM PCR) for bacterial strain differentiation
Pulsed Field Gel Electrophoresis (EK-PFGE), Southern blot hybridization with the midrepeat sequence Ca3, PCR-Restriction Fragment Length Polymorphism (RFLP), macrorestriction analysis of genomic DNA followed by pulsed field gel electrophoresis (REA-PFGE), Amplified
A total of 123 strains of C. albicans were typed with the REA-PFGE method
Summary
We have previously reported the use of PCR Melting Profile (PCR MP) technique based on using low denaturation temperatures during ligation mediated PCR (LM PCR) for bacterial strain differentiation. The aim of the current study was to evaluate this method for intraspecies differentiation of Candida albicans strains. It is still generally accepted that Candida albicans is the major etiologic pathogen of candidiasis (candidemia, candidosis). Over the last several years, molecular methods have proven quite useful to study strain relatedness, allowing retrospective examinations of putative candidosis outbreaks and assessments of the epidemiological aspects of such outbreaks. Microsatellite Locus Analysis (PMLA), analysis of the repetive sequences (RPSs) and RAPD method have all (page number not for citation purposes) been used for strain typing of C. albicans [3,4,5,6,7,8,9,10,11,12,13]. The method is highly reproducible, and data can be archived in Web-based databases accessible to all users
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