PMID- 16081953
OWN - NLM
STAT- MEDLINE
DCOM- 20050915
LR  - 20181113
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 43
IP  - 8
DP  - 2005 Aug
TI  - Superiority of molecular techniques for identification of gram-negative, 
      oxidase-positive rods, including morphologically nontypical Pseudomonas 
      aeruginosa, from patients with cystic fibrosis.
PG  - 4070-5
AB  - Phenotypic identification of gram-negative bacteria from Cystic Fibrosis (CF) 
      patients carries a high risk of misidentification. Therefore, we compared the 
      results of biochemical identification by API 20NE with 16S rRNA gene sequencing 
      in 88 gram-negative, oxidase-positive rods, other than morphologically and 
      biochemically typical P. aeruginosa, from respiratory secretions of CF patients. 
      The API 20NE allowed correct identification of the bacterial species in 15 out of 
      88 (17%) isolates investigated. Agreement between the API and the 16S rRNA gene 
      sequencing results was high only in isolates with an API result classified as 
      "excellent identification". Even API results classified as "very good 
      identification" or "good identification" showed a high rate of misidentification 
      (67% and 84%). Fifty-two isolates of morphological and biochemical nontypical 
      Pseudomonas aeruginosa, representing 59% of all isolates investigated, were not 
      identifiable or misidentified in the API 20NE. Therefore, rapid molecular 
      diagnostic techniques like real-time PCR and fluorescence in situ hybridization 
      (FISH) were evaluated in this particular group of bacteria for identification of 
      the clinically most relevant pathogen, P. aeruginosa. The LightCycler PCR assay 
      with a P. aeruginosa-specific probe showed a sensitivity and specificity of 98.1% 
      and 100%, respectively. For FISH analysis, a newly designed P. 
      aeruginosa-specific probe had a sensitivity and specificity of 100%. In 
      conclusion, molecular methods are superior over biochemical tests for 
      identification of gram-negative, oxidase-positive rods in CF patients. In 
      addition, real-time PCR and FISH allowed identification of morphologically 
      nontypical isolates of P. aeruginosa within a few hours.
FAU - Wellinghausen, Nele
AU  - Wellinghausen N
AD  - Department of Medical Microbiology and Hygiene, University of Ulm, 
      Robert-Koch-Str. 8, D-89081 Ulm, Germany. nele.wellinghausen@medizin.uni-ulm.de
FAU - Kothe, Juliane
AU  - Kothe J
FAU - Wirths, Beate
AU  - Wirths B
FAU - Sigge, Anja
AU  - Sigge A
FAU - Poppert, Sven
AU  - Poppert S
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (RNA, Ribosomal, 16S)
RN  - 0 (Reagent Strips)
RN  - EC 1.- (Oxidoreductases)
SB  - IM
MH  - Cystic Fibrosis/*microbiology
MH  - Gram-Negative Bacteria/enzymology/genetics/*isolation & purification
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Oxidoreductases/*analysis
MH  - Polymerase Chain Reaction
MH  - Pseudomonas aeruginosa/enzymology/genetics/*isolation & purification
MH  - RNA, Ribosomal, 16S/genetics
MH  - Reagent Strips
MH  - Sequence Analysis, DNA
PMC - PMC1233906
EDAT- 2005/08/06 09:00
MHDA- 2005/09/16 09:00
PMCR- 2006/02/01
CRDT- 2005/08/06 09:00
PHST- 2005/08/06 09:00 [pubmed]
PHST- 2005/09/16 09:00 [medline]
PHST- 2005/08/06 09:00 [entrez]
PHST- 2006/02/01 00:00 [pmc-release]
AID - 43/8/4070 [pii]
AID - 0203-05 [pii]
AID - 10.1128/JCM.43.8.4070-4075.2005 [doi]
PST - ppublish
SO  - J Clin Microbiol. 2005 Aug;43(8):4070-5. doi: 10.1128/JCM.43.8.4070-4075.2005.