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  • Spectrophotometric reading of EUCAST antifungal susceptibility testing of Aspergillus fumigatus.

Spectrophotometric reading of EUCAST antifungal susceptibility testing of Aspergillus fumigatus.

Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases (2016-10-30)
J Meletiadis, K Leth Mortensen, P E Verweij, J W Mouton, M C Arendrup
ABSTRACT

Given the increasing number of antifungal drugs and the emergence of resistant Aspergillus isolates, objective, automated and high-throughput antifungal susceptibility testing is important. The EUCAST E.Def 9.3 reference method for MIC determination of Aspergillus species relies on visual reading. Spectrophotometric reading was not adopted because of concern that non-uniform filamentous growth might lead to unreliable and non-reproducible results. We therefore evaluated spectrophotometric reading for the determination of MICs of antifungal azoles against Aspergillus fumigatus. Eighty-eight clinical isolates of A. fumigatus were tested against four medical azoles (posaconazole, voriconazole, itraconazole, isavuconazole) and one agricultural azole (tebuconazole) with EUCAST E.Def 9.3. The visually determined MICs (complete inhibition of growth) were compared with spectrophotometrically determined MICs and essential (±1 twofold dilution) and categorical (susceptible/intermediate/resistant or wild-type/non-wild-type) agreement was calculated. Spectrophotometric data were analysed with regression analysis using the E Using the 5% cut-off, high essential (92%-97%) and categorical (93%-99%) agreement (<6% errors) was found between spectrophotometric and visual MICs. The EC Spectrophotometric determination of MICs of antifungal drugs may increase objectivity, and allow automation and high-throughput of EUCAST E.Def 9.3 antifungal susceptibility testing of Aspergillus species.

MATERIALS
Product Number
Brand
Product Description

Supelco
Tebuconazole, PESTANAL®, analytical standard
Supelco
Posaconazole, VETRANAL®, analytical standard