Relation of the polymorphism of cyp51A sequence and the susceptibility of Aspergillus fumigatus  isolates to triazoles determinated by commercial gradient test (Etest) and by reference methods

Urszula Nawrot; Beata Sulik-Tyszka; Ewelina Kurzyk; Katarzyna Wlodarczyk; Marta Wróblewska; Grzegorz Władysław Basak; Anna Brillowska-Dąbrowska

doi:10.18388/abp.2017_1571

Urszula Nawrot Department of Pharmaceutical Microbiology and Parasitology Wroclaw Medical University
Beata Sulik-Tyszka Department of Microbiology, Central Clinical Hospital, Warsaw Medical University, Warsaw, Poland
Ewelina Kurzyk Department of Molecular Biotechnology and Microbiology, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, Poland
Katarzyna Wlodarczyk Department of Pharmaceutical Microbiology and Parasitology, Faculty of Pharmacy, Wrocław Medical University, Borowska 213, 50-566 Wrocław, Poland
Marta Wróblewska Department of Microbiology, Central Clinical Hospital, Warsaw Medical University, Warsaw, Poland Department of Dental Microbiology, Warsaw Medical University, Warsaw, Poland
Grzegorz Władysław Basak Department of Haematology, Oncology and Internal Medicine, Medical University of Warsaw, Warsaw, Poland
Anna Brillowska-Dąbrowska Department of Molecular Biotechnology and Microbiology, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, Poland

DOI: https://doi.org/10.18388/abp.2017_1571

Keywords: Aspergillus fumigatus, triazole resistance, susceptibility testing, Etest, cyp51A sequence

Abstract

The aim of this study was to evaluate the accuracy of commercial gradient test (Etest) in the detection of triazole resistant Aspergillus fumigatus isolates using reference microdilution methods and the analysis of sequences of the cyp 51A gene. The study was performed on twenty clinical isolates which were identified as Aspergillus fumigatus on the base of the DNA sequences of the ITS1-2 fragment of ribosomal DNA and the β-tubulin gene, out of them seventeen isolates showed wild -type of the cyp51A sequence and three were positive for the mutation TR34/L98H . All isolates were tested for the susceptibility to itraconazole (ITZ), voriconazole (VOR) and posaconasole (POS) using microdilution methods, according to EUCAST and CLSI protocols, as well as Etest. The results of microdilution and Etests were analysed separately according to clinical breakpoints (CBP) defined by EUCAST version 7.0 and epidemiological cut off values (ECV). Etest as well as reference methods excellently recognised the WT isolates, which were susceptible to all tested triazoles, regardless of the method and CBP or ECV criteria used. The Etest recognized three non-WT isolates as resistant or intermediately sensitive to ITZ and POS and one as resistant to VOR. The categorical concordance between Etests and EUCAST and Etests and the CLSI method ranged from 90 to 100%.

The interpretation of results of routine A. fumigatus Etests require great caution. The use of the confirmative examinations with reference AST methods as well as with molecular tests are recommended.

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