Real-Time PCR Identification of Six Malassezia Species.
Lipophilic yeast Malassezia species is widely found on the skin surface of humans and other animals. This fungus can cause pityriasis versicolor, Malassezia folliculitis, and seborrheic dermatitis. Still now, there is a problem with species identification of Malassezia with conventional methods. We developed a real-time polymerase chain reaction (PCR) assay with multiple hybridization probes for detecting M. globosa, M. furfur, M. restricta, M. sympodialis, M. slooffiae, and M. pachydermatis. The amplification curves and specific melting peaks of the probes hybridized with real-time PCR product were used for species identifications. The assay was further evaluated on 120 samples which were performed by swabbing from 60 domestic animals (23 goats, 10 dogs, 15 cows, 3 cats, 8 rabbits, and 1 donkey) and in 70 human samples (28 patients with pityriasis versicolor, 17 breeders, and 25 control group). Fifteen M. pachydermatis were identified from animals. From human, 61 isolates were identified as M. globosa (28), M. furfur (15), M. restricta (6), M. sympodialis (8), M. slooffiae (2), and M. pachydermatis (2). Eight cases of co-detection from 6 patients and 2 breeders were revealed. Our findings show that the assay was highly effective in identifying Malassezia species. The application of multiplex real-time PCR provides a sensitive and rapid identification system for Malassezia species, which may be applied in further epidemiological surveys from clinical samples.
Fungal and Parasitic Molecular Biology Laboratory, Faculty of Medicine, University of Sfax, Rue Magida Boulila, Sfax, 3029, Tunisia.
Amin
A
Ilahi
Fungal and Parasitic Molecular Biology Laboratory, Faculty of Medicine, University of Sfax, Rue Magida Boulila, Sfax, 3029, Tunisia. ineshadrich@yahoo.fr.
Inès
I
Hadrich
Fungal and Parasitic Molecular Biology Laboratory, Faculty of Medicine, University of Sfax, Rue Magida Boulila, Sfax, 3029, Tunisia.
Sourour
S
Neji
Fungal and Parasitic Molecular Biology Laboratory, Faculty of Medicine, University of Sfax, Rue Magida Boulila, Sfax, 3029, Tunisia.
Houaida
H
Trabelsi
Fungal and Parasitic Molecular Biology Laboratory, Faculty of Medicine, University of Sfax, Rue Magida Boulila, Sfax, 3029, Tunisia.
Fattouma
F
Makni
Fungal and Parasitic Molecular Biology Laboratory, Faculty of Medicine, University of Sfax, Rue Magida Boulila, Sfax, 3029, Tunisia.
Ali
A
Ayadi
D000818:Animals; D002415:Cats; D002417:Cattle / Q000382:microbiology; D004285:Dogs; D018397:Equidae / Q000382:microbiology; D006041:Goats / Q000382:microbiology; D006801:Humans; D008290:Malassezia / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D058889:Molecular Typing / Q000379:methods*; D016533:Mycological Typing Techniques / Q000379:methods*; D009691:Nucleic Acid Denaturation / Q000235:genetics; D012335:RNA, Ribosomal / Q000235:genetics; D011817:Rabbits; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012867:Skin / Q000382:microbiology; D014010:Tinea Versicolor / Q000382:microbiology
D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S
Domestic animals; Malassezia; Molecular identification; PCR; Probes