POL Scientific / JBM / Volume 3 / Issue 4 / DOI: 10.14440/jbm.2016.125
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Performance of pyrosequencing versus MALDI-TOF MS in bacteria identification in chronic lung disease

Lucie Navrátilová1,2 Petra Procházková1 Jan Bardoň1,3 Radko Novotný1 Martin Zápalka4 Petr Jakubec5 Jaromír Zatlou-kal5 Vítězslav Kolek5 František Kopřiva4 Pavla Flodrová7 Vladislav Raclavský1,6
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1 Department of Microbiology, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hnevotinska 3, 779 00 Olomouc, Czech Republic
2 Laboratory of Growth Regulators, Palacký University Olomouc & Institute of Experimental Botany AS CR, Slechtitelu 27, 783 71 Olomouc, Czech Republic
3 State Veterinary Institute Olomouc, Jakoubka ze Stribra 1, 779 00 Olomouc, Czech Republic
4 Department of Pediatrics, University Hospital Olomouc, I. P. Pavlova 6, Olomouc, Czech Republic
5 Department of Respiratory Medicine, University Hospital Olomouc, I. P. Pavlova 6, Olomouc, Czech Republic
6 Institute of Molecular and Translational Medicine, Hnevotinska 5, Olomouc, Czech Republic
7 Department of Clinical and Molecular Pathology, Faculty of Medicine and Dentistry, Palacký University Olomouc, Hnevotinska 3, 779 00 Olomouc, Czech Republic
JBM 2016 , 3(4), 1;
Published: 13 August 2016
© 2016 by the author. Licensee POL Scientific, USA. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

Rapid identification of the etiological agent in bacterial infection is necessary for correct diagnosis and appropriate therapy. In general, identification of pure cultures of bacteria using conventional phenotyping techniques requires 4-24 hours. Recently available new molecular technologies offer the potential of same day species identification once pure culture is available. Our aim was to evaluate the performance of rDNA V1 hypervariable region pyrosequencing, and the whole cell MALDI-TOF MS protein profiling in routine species identification. During the period from June 2012 to June 2014, 1.140 pure culture isolates were recovered from 402 samples from 126 patients suffering cystic fibrosis, chronic obstructive pulmonary disease or bronchiectasis. All the isolates were subjected to species identification by both techniques. Unfortunately, pyrosequencing was able to reach the species level in 43.2% of isolates only, whereas MALDI-TOF was clearly superior with 96.8% respectively. The overall sensitivity values also clearly underlined the superiority of MALDI-TOF MS with 96.8% compared to 85.1% achieved by pyrosequencing. Generally, MALDI-TOF MS turned out to be the best suitable technique in routine bacterial identification, whereas pyrosequencing could be recommended as the method of choice particularly in situations where MALDI-TOF MS fails to identify rare species.

Keywords
MALDI-TOF MS
pyrosequencing
16S rDNA
Cystic fibrosis
Chronic obstructive pulmonary disease
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Journal of Biological Methods, Electronic ISSN: 2326-9901 Print ISSN: TAB, Published by POL Scientific