Identification and differentiation of clinical isolates using PCR-RFLP and RAPD methods

A. Hryncewicz-Gwóźdź (1) , T. Jagielski (2) , A. Dobrowolska (3) , J. C. Szepietowski (1) , E. Baran (1)

1 Department of Dermatology, Venereology and Allergology
2 Department of Applied Microbiology [Warsaw]
3 Department of Genetics of Microorganisms

represents the most frequently isolated causative agent of superficial dermatophyte infections. Several genotyping methods have recently been introduced to improve the delineation between pathogenic fungi at both the species and the strain levels. The purpose of this study was to apply selected DNA fingerprinting methods to the identification and strain discrimination of clinical isolates. Fifty-seven isolates from as many tinea patients were subjected to species identification by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis and strain differentiation using a randomly amplified polymorphic DNA (RAPD) method, with two primers designated 1 and 6. Using PCR-RFLP, 55 of the isolates studied were confirmed to be . Among those, a total of 40 and five distinct profiles were obtained by RAPD with primers 1 and 6, respectively. The combination of profiles from both RAPD assays resulted in 47 genotypes and an overall genotypic diversity rate of 85.4%. A dendrogram analysis performed on the profiles generated by RAPD with primer 1 showed most of the isolates (87.3%) to be genetically related. PCR-RFLP serves as a rapid and reliable method for the identification of species, while the RAPD analysis is rather a disadvantageous tool for strain typing.

Mots clés: Life Sciences

Domaines: Maladies infectieuses

A. Hryncewicz-Gwóźdź, T. Jagielski, A. Dobrowolska, J. C. Szepietowski, E. Baran. Identification and differentiation of clinical isolates using PCR-RFLP and RAPD methods. European Journal of Clinical Microbiology and Infectious Diseases, 2011, 30 (6), pp.727-731. ⟨10.1007/s10096-010-1144-3⟩. ⟨hal-00680180⟩