Download Diagnosis of human pathogenic fungi by molecular and MALDI

Diagnosis of human pathogenic fungi by molecular and
MALDI-ToF methods
Ali Abdul Hussein S. AL-Janabi and Sybren de Hoog
CBS-KNAW Fungal Biodiversity Centre, Utrecht, The Netherlands
Dept. of Clinical laboratories, College of Applied Medical Sciences, University of
Kerbala, Iraq
[email protected]
Abstract
There are many different species of fungi caused human infections. Human skin are
infected by various genera which are common distributed all over the world.
Dermatophytes and Hortaea werineckii are known to cause human skin infection
called Tinea. The name of Tinea caused y dermatophytes depends on the location of
disease in skin, while Tinea caused by Hortaea werineckii is called Tinea Niger. Two
methods were used to diagnosis of those skin pathogenic fungi, MALDI-ToF and
Molecular methods. All of fungal strains were obtained from the CBS culture
collection (Netherlands). Total of 300 strains of dermatophytes and 45 strains of
Hortaea werineckii were re-identified. MALDI-ToF as new technique for diagnosis
of fungi was performed in BioMerieux instrument. Three genes were chosen to reidentification of Hortaea werineckii strains. Internal transcribed spacer regions (ITS),
translation elongation factor-1 alpha (EF1α), and partial β- tubuln (β-Tubulin) gene
were amplified with primer pairs (ITS1& ITS4), (EF1-983F & EF1-2218R) and (T1F
& Bt2bR) respectively. The sequences were aligned by Bionumerics computer
software for Phylogenic tree of Hortaea werineckii. The results of MALDI-ToF were
confirmed the fungi identified by CBS research center which means the efficiency of
this method to use for fungal diagnosis. Gene analysis for Hortaea werineckii showed
a close relationship between strains in phylogenic tree with exception of three far
strains: Teratosphaeria hortaea CBS 124156, Hortaea thailandica CBS 125423, and
Hortaea acidophila CBS 113389.