Identification of Malassezia species from patient skin scales by PCR-RFLP
Φόρτωση...
Ημερομηνία
Συγγραφείς
Gaitanis, G.
Velegraki, A.
Frangoulis, E.
Mitroussia, A.
Tsigonia, A.
Tzimogianni, A.
Katsambas, A.
Legakis, N. J.
Τίτλος Εφημερίδας
Περιοδικό ISSN
Τίτλος τόμου
Εκδότης
Περίληψη
Τύπος
Είδος δημοσίευσης σε συνέδριο
Είδος περιοδικού
peer-reviewed
Είδος εκπαιδευτικού υλικού
Όνομα συνεδρίου
Όνομα περιοδικού
Clin Microbiol Infect
Όνομα βιβλίου
Σειρά βιβλίου
Έκδοση βιβλίου
Συμπληρωματικός/δευτερεύων τίτλος
Περιγραφή
OBJECTIVE: This study was aimed at the development of a DNA-based procedure directly applicable to pathological skin scales and at the assessment of its value in rapid laboratory confirmation and identification of each of the seven Malassezia species. These lipophilic basidiomycetous yeasts in predisposed individuals are involved in pityriasis versicolor, seborrheic dermatitis, blepharitis, folliculitis, atopic dermatitis and fungemia. Standard identification procedures to species level are available, but so far no system for direct detection and characterization of Malassezia species in clinical specimens is available. METHODS: Malassezia DNA was extracted from pathological skin scales by a modified hexadecyltrimethylammonium bromide (CTAB) method and amplified by single and nested polymerase chain reaction (PCR), assays using the general fungal ITS 1/4 and 3/4 primers for amplification of sequences from the Malassezia major ribosomal DNA complex. Restriction fragment length polymorphism (RFLP) analysis of PCR products was used in subsequent species identification. DNA extracted from culture-positive skin scales was also tested by PCR and the RFLP patterns obtained were analyzed. RESULTS: A total of 36 isolates were tested. Distinct pure culture and skin-scale ITS 3/4 HinfI and AluI restriction patterns differentially identified M. furfur, M. globosa, M. restricta, M. sympodialis, M. pachydermatis, M. obtusa and M. slooffiae. Malassezia DNA was extracted from pathological skin scales and RFLP identified solitary and multiple Malassezia species in the same specimen. Molecular identification was confirmed by cultures and biochemical tests. Concurrent detection and identification of Candida and Yarrowia species was also feasible from skin scales. CONCLUSION: The proposed method, described for the first time, could provide a sensitive and rapid detection and identification system for Malassezia species, which may be applied to epidemiological surveys and routine practice.
Περιγραφή
Λέξεις-κλειδιά
Animals, Dermatitis, Seborrheic/microbiology, Dogs, Genes, Bacterial/genetics, Humans, Malassezia/*classification/genetics/*isolation & purification, Polymerase Chain Reaction/*methods, *Polymorphism, Restriction Fragment Length, Reproducibility of Results, Sensitivity and Specificity, Skin Diseases/*microbiology/*pathology, Tinea Versicolor/microbiology
Θεματική κατηγορία
Παραπομπή
Σύνδεσμος
http://www.ncbi.nlm.nih.gov/pubmed/12010171
http://onlinelibrary.wiley.com/store/10.1046/j.1469-0691.2002.00383.x/asset/j.1469-0691.2002.00383.x.pdf?v=1&t=h0ta0j3q&s=dc7200028b5e422939f7b727c76756573221cb5d
http://onlinelibrary.wiley.com/store/10.1046/j.1469-0691.2002.00383.x/asset/j.1469-0691.2002.00383.x.pdf?v=1&t=h0ta0j3q&s=dc7200028b5e422939f7b727c76756573221cb5d
Γλώσσα
en
Εκδίδον τμήμα/τομέας
Όνομα επιβλέποντος
Εξεταστική επιτροπή
Γενική Περιγραφή / Σχόλια
Ίδρυμα και Σχολή/Τμήμα του υποβάλλοντος
Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής