PMID- 23646201
OWN - NLM
STAT- MEDLINE
DCOM- 20131126
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 4
DP  - 2013
TI  - Rapid discrimination of Haemophilus influenzae, H. parainfluenzae, and H. 
      haemolyticus by fluorescence in situ hybridization (FISH) and two matrix-assisted 
      laser-desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS) 
      platforms.
PG  - e63222
LID - 10.1371/journal.pone.0063222 [doi]
LID - e63222
AB  - BACKGROUND: Due to considerable differences in pathogenicity, Haemophilus 
      influenzae, H. parainfluenzae and H. haemolyticus have to be reliably 
      discriminated in routine diagnostics. Retrospective analyses suggest frequent 
      misidentifications of commensal H. haemolyticus as H. influenzae. In a 
      multi-center approach, we assessed the suitability of fluorescence in situ 
      hybridization (FISH) and matrix-assisted laser-desorption-ionization 
      time-of-flight mass-spectrometry (MALDI-TOF-MS) for the identification of H. 
      influenzae, H. parainfluenzae and H. haemolyticus to species level. METHODOLOGY: 
      A strain collection of 84 Haemophilus spp. comprising 50 H. influenzae, 25 H. 
      parainfluenzae, 7 H. haemolyticus, and 2 H. parahaemolyticus including 77 
      clinical isolates was analyzed by FISH with newly designed DNA probes, and two 
      different MALDI-TOF-MS systems (Bruker, Shimadzu) with and without prior formic 
      acid extraction. PRINCIPAL FINDINGS: Among the 84 Haemophilus strains analyzed, 
      FISH led to 71 correct results (85%), 13 uninterpretable results (15%), and no 
      misidentifications. Shimadzu MALDI-TOF-MS resulted in 59 correct identifications 
      (70%), 19 uninterpretable results (23%), and 6 misidentifications (7%), using 
      colony material applied directly. Bruker MALDI-TOF-MS with prior formic acid 
      extraction led to 74 correct results (88%), 4 uninterpretable results (5%) and 6 
      misidentifications (7%). The Bruker MALDI-TOF-MS misidentifications could be 
      resolved by the addition of a suitable H. haemolyticus reference spectrum to the 
      system's database. In conclusion, no analyzed diagnostic procedure was free of 
      errors. Diagnostic results have to be interpreted carefully and alternative tests 
      should be applied in case of ambiguous test results on isolates from seriously 
      ill patients.
FAU - Frickmann, Hagen
AU  - Frickmann H
AD  - Department of Tropical Medicine at the Bernhard Nocht Institute, German Armed 
      Forces Hospital of Hamburg, Hamburg, Germany. Frickmann@bni-hamburg.de
FAU - Christner, Martin
AU  - Christner M
FAU - Donat, Martina
AU  - Donat M
FAU - Berger, Anja
AU  - Berger A
FAU - Essig, Andreas
AU  - Essig A
FAU - Podbielski, Andreas
AU  - Podbielski A
FAU - Hagen, Ralf Matthias
AU  - Hagen RM
FAU - Poppert, Sven
AU  - Poppert S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20130430
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
SB  - IM
MH  - Haemophilus/*chemistry/classification/*genetics/isolation & purification
MH  - Haemophilus Infections/diagnosis
MH  - Haemophilus influenzae/genetics
MH  - Haemophilus parainfluenzae/genetics
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - Retrospective Studies
MH  - *Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PMC - PMC3639997
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2013/05/07 06:00
MHDA- 2013/12/16 06:00
PMCR- 2013/04/30
CRDT- 2013/05/07 06:00
PHST- 2012/10/17 00:00 [received]
PHST- 2013/04/01 00:00 [accepted]
PHST- 2013/05/07 06:00 [entrez]
PHST- 2013/05/07 06:00 [pubmed]
PHST- 2013/12/16 06:00 [medline]
PHST- 2013/04/30 00:00 [pmc-release]
AID - PONE-D-12-32252 [pii]
AID - 10.1371/journal.pone.0063222 [doi]
PST - epublish
SO  - PLoS One. 2013 Apr 30;8(4):e63222. doi: 10.1371/journal.pone.0063222. Print 2013.