Altay Atalay1, Ayse Nedret Koc2, Ahmet Suel3, Hafize Sav2, Gonca Demir2, Ferhan Elmali4, Nuri Cakir2, Seyedmojtaba Seyedmousavi5,6,7. 1. Faculty of Medicine, Department of Medical Microbiology, University of Erciyes, Kayseri, Turkey. altayatalay@gmail.com. 2. Faculty of Medicine, Department of Medical Microbiology, University of Erciyes, Kayseri, Turkey. 3. BioMérieux Diagnostic, Inc, Istanbul, Turkey. 4. Faculty of Medicine, Department of Biostatistics, University of Erciyes, Kayseri, Turkey. 5. Department of Medical Microbiology, Radboud University Medical Centre, Nijmegen, The Netherlands. 6. Department of Medical Microbiology and Infectious Diseases, ErasmusMC, The Netherlands. 7. Invasive Fungi Research Center, Mazandaran University of Medical Sciences, Sari, Iran.
Abstract
BACKGROUND: Aspergillus species cause a wide range of diseases in humans, including allergies, localized infections, or fatal disseminated diseases. Rapid detection and identification of Aspergillus spp. facilitate effective patient management. In the current study we compared conventional morphological methods with PCR sequencing, rep-PCR, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for the identification of Aspergillus strains. MATERIALS AND METHODS: A total of 24 consecutive clinical isolates of Aspergillus were collected during 2012-2014. Conventional morphology and rep-PCR were performed in our Mycology Laboratory. The identification, evaluation, and reporting of strains using MALDI-TOF-MS were performed by BioMérieux Diagnostic, Inc. in Istanbul. DNA sequence analysis of the clinical isolates was performed by the BMLabosis laboratory in Ankara. RESULTS: Samples consisted of 18 (75%) lower respiratory tract specimens, 3 otomycosis (12.5%) ear tissues, 1 sample from keratitis, and 1 sample from a cutaneous wound. According to DNA sequence analysis, 12 (50%) specimens were identified as A. fumigatus, 8 (33.3%) as A. flavus, 3 (12.5%) as A. niger, and 1 (4.2%) as A. terreus. Statistically, there was good agreement between the conventional morphology and rep-PCR and MALDI-TOF methods; kappa values were κ = 0.869, 0.871, and 0.916, respectively (P < 0.001). CONCLUSION: The good level of agreement between the methods included in the present study and sequence method could be due to the identification of Aspergillus strains that were commonly encountered. Therefore, it was concluded that studies conducted with a higher number of isolates, which include other Aspergillus strains, are required.
BACKGROUND:Aspergillus species cause a wide range of diseases in humans, including allergies, localized infections, or fatal disseminated diseases. Rapid detection and identification of Aspergillus spp. facilitate effective patient management. In the current study we compared conventional morphological methods with PCR sequencing, rep-PCR, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for the identification of Aspergillus strains. MATERIALS AND METHODS: A total of 24 consecutive clinical isolates of Aspergillus were collected during 2012-2014. Conventional morphology and rep-PCR were performed in our Mycology Laboratory. The identification, evaluation, and reporting of strains using MALDI-TOF-MS were performed by BioMérieux Diagnostic, Inc. in Istanbul. DNA sequence analysis of the clinical isolates was performed by the BMLabosis laboratory in Ankara. RESULTS: Samples consisted of 18 (75%) lower respiratory tract specimens, 3 otomycosis (12.5%) ear tissues, 1 sample from keratitis, and 1 sample from a cutaneous wound. According to DNA sequence analysis, 12 (50%) specimens were identified as A. fumigatus, 8 (33.3%) as A. flavus, 3 (12.5%) as A. niger, and 1 (4.2%) as A. terreus. Statistically, there was good agreement between the conventional morphology and rep-PCR and MALDI-TOF methods; kappa values were κ = 0.869, 0.871, and 0.916, respectively (P < 0.001). CONCLUSION: The good level of agreement between the methods included in the present study and sequence method could be due to the identification of Aspergillus strains that were commonly encountered. Therefore, it was concluded that studies conducted with a higher number of isolates, which include other Aspergillus strains, are required.
Authors: Justin M Hettick; Brett J Green; Amanda D Buskirk; Michael L Kashon; James E Slaven; Erika Janotka; Francoise M Blachere; Detlef Schmechel; Donald H Beezhold Journal: Anal Biochem Date: 2008-06-05 Impact factor: 3.365
Authors: William J Steinbach; Daniel K Benjamin; Dimitrios P Kontoyiannis; John R Perfect; Irja Lutsar; Kieren A Marr; Michail S Lionakis; Harrys A Torres; Hasan Jafri; Thomas J Walsh Journal: Clin Infect Dis Date: 2004-06-28 Impact factor: 9.079