Literature DB >> 26926455

Conventional Morphology Versus PCR Sequencing, rep-PCR, and MALDI-TOF-MS for Identification of Clinical Aspergillus Isolates Collected Over a 2-Year Period in a University Hospital at Kayseri, Turkey.

Altay Atalay1, Ayse Nedret Koc2, Ahmet Suel3, Hafize Sav2, Gonca Demir2, Ferhan Elmali4, Nuri Cakir2, Seyedmojtaba Seyedmousavi5,6,7.   

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.
© 2016 Wiley Periodicals, Inc.

Entities:  

Keywords:  Aspergillus; MALDI-TOF-MS; rep-PCR; sequencing

Mesh:

Year:  2016        PMID: 26926455      PMCID: PMC6807001          DOI: 10.1002/jcla.21932

Source DB:  PubMed          Journal:  J Clin Lab Anal        ISSN: 0887-8013            Impact factor:   2.352


  32 in total

1.  Use of the Diversi Lab System for species and strain differentiation of Fusarium species isolates.

Authors:  M Healy; K Reece; D Walton; J Huong; S Frye; I I Raad; D P Kontoyiannis
Journal:  J Clin Microbiol       Date:  2005-10       Impact factor: 5.948

2.  MALDI-TOF mass spectrometry identification of filamentous fungi in the clinical laboratory.

Authors:  Stéphane Ranque; Anne-Cécile Normand; Carole Cassagne; Jean-Benjamin Murat; Nathalie Bourgeois; Frédéric Dalle; Martine Gari-Toussaint; Patrick Fourquet; Marijke Hendrickx; Renaud Piarroux
Journal:  Mycoses       Date:  2013-07-29       Impact factor: 4.377

3.  Fungal infection of the ear: a common problem in the north eastern part of Haryana.

Authors:  K R Aneja; Chetan Sharma; Radhika Joshi
Journal:  Int J Pediatr Otorhinolaryngol       Date:  2010-03-26       Impact factor: 1.675

4.  Use of the Bruker MALDI Biotyper for identification of molds in the clinical mycology laboratory.

Authors:  Bettina Schulthess; Raphael Ledermann; Forouhar Mouttet; Andrea Zbinden; Guido V Bloemberg; Erik C Böttger; Michael Hombach
Journal:  J Clin Microbiol       Date:  2014-05-21       Impact factor: 5.948

5.  Repetitive-sequence-PCR-based DNA fingerprinting using the Diversilab system for identification of commonly encountered dermatophytes.

Authors:  June I Pounder; Sheri Williams; Dewey Hansen; Mimi Healy; Kristy Reece; Gail L Woods
Journal:  J Clin Microbiol       Date:  2005-05       Impact factor: 5.948

Review 6.  Identification of fungal microorganisms by MALDI-TOF mass spectrometry.

Authors:  Jana Chalupová; Martin Raus; Michaela Sedlářová; Marek Sebela
Journal:  Biotechnol Adv       Date:  2013-11-07       Impact factor: 14.227

7.  Use of a rep-PCR system to predict species in the Aspergillus section Nigri.

Authors:  Edwin R Palencia; Maren A Klich; Anthony E Glenn; Charles W Bacon
Journal:  J Microbiol Methods       Date:  2009-07-21       Impact factor: 2.363

8.  Discrimination of Aspergillus isolates at the species and strain level by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry fingerprinting.

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

Review 9.  Aspergillus flavus: an emerging non-fumigatus Aspergillus species of significance.

Authors:  Suganthini Krishnan; Elias K Manavathu; Pranatharthi H Chandrasekar
Journal:  Mycoses       Date:  2009-01-14       Impact factor: 4.377

10.  Infections due to Aspergillus terreus: a multicenter retrospective analysis of 83 cases.

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

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  2 in total

Review 1.  Mass spectrometry in research laboratories and clinical diagnostic: a new era in medical mycology.

Authors:  Hasti Kamali Sarvestani; Alireza Ramandi; Muhammad Ibrahim Getso; Taraneh Razavyoon; Javad Javidnia; Miaad Banay Golrizi; Ali-Akbar Saboor-Yaraghi; Saham Ansari
Journal:  Braz J Microbiol       Date:  2022-03-28       Impact factor: 2.214

Review 2.  Mycotic Keratitis-A Global Threat from the Filamentous Fungi.

Authors:  Jeremy J Hoffman; Matthew J Burton; Astrid Leck
Journal:  J Fungi (Basel)       Date:  2021-04-03
  2 in total

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