Investigation Of Primary Clinical Samples Of Immunocompromised Patients, Azole Resistant Clinical And Environmental Isolates For Aspergillus Fumigatus Cyp51A Key Mutations Using Sensitive PCR Assays And Consecutive DNA Sequence Analysis

Ref ID: 18705

Author:

B. Spiess, PhD (Doctor of Philosophy) – Scientist1, M. Reinwald, MD – Resident 1, S. J. Howard, PhD – Scientist 2, U. Gross, MD – Professor 3, O. Bader, PhD – Scientist 3, P. M. Rath, MD – Professor 4, W. Seifarth, PhD – Scientist 1, S. Will, TA – Re

Author address:

1Mannheim Univ. Hosp., 3rd Dept. of Internal Med., Mannheim, Germany, 2Univ. of Liverpool, Translational Med., Liverpool, United Kingdom, 3Inst. of Med. Microbiol., Göttingen Univ. Hosp., Göttingen, Germany, 4Inst. of Med. Microbiol., Essen Univ. Hos

Full conference title:

52nd Annual ICAAC

Date: 9 September 2014

Abstract:

Background: The incidence of azole resistance in Aspergillus fumigatus is rising. As the diagnosis of invasive aspergillosis is rarely based on positive culture yield in immunocompromised patients, detection of azole resistance directly from clinical samples and isolates would be highly valuable.
Methods: We screened 61 primary clinical samples (BAL and biopsies) from 58 immunocompromised patients for cyp51A TR (tandem repeat), L98H and M220 gene alterations using sensitive PCR assays and consecutive DNA sequence analysis. Furthermore PCR and sequence analysis of 3 azole resistant clinical isolates and 3 environmental strains was performed.
Results: DNA sequence analysis of primary clinical samples revealed a single L98H mutation in a lung tissue specimen of a COPD patient and a L98H alteration in combination with the TR in a brain tissue sample of a patient with acute lymphoblastic leukemia. Investigation of three azole resistant strains of A. fumigatus in clinical isolates revealed the TR and L98H alterations in two cases, additionally a M220T alteration in the third case. Sequence analysis of one environmental strain revealed both the L98H and TR alterations, in the second strain a M220V alteration.
Conclusions: Our results using clinical samples and azole resistant A. fumigatus clinical and environmental strains show the feasibility of the approach. We consider our assay of high epidemiological and clinical relevance to detect azole resistance both in clinical and environmental samples.

Abstract Number: M-335

Conference Year: 2012

Link to conference website: NULL

New link: NULL

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