Identification of Bacteria by Polymerase Chain Reaction Followed by Liquid Chromatography−Mass Spectrometry

Bloodstream infections are an important cause of serious morbidity and mortality. Rapid detection and identification of specific pathogens from blood or other clinical specimens could improve the rational use of antimicrobial therapy in clinical medicine and have a great impact on the outcome of pat...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2005-07, Vol.77 (14), p.4563-4570
Main Authors: Mayr, Bettina M, Kobold, Uwe, Moczko, Martin, Nyeki, Agnes, Koch, Thomas, Huber, Christian G
Format: Article
Language:English
Subjects:
Analytical chemistry
Bacteria
Bacteria - classification
Bacteria - isolation & purification
Base Sequence
Biological and medical sciences
Biotechnology
Chemistry
Chromatographic methods and physical methods associated with chromatography
Chromatography
Chromatography, Liquid - methods
DNA polymerase
DNA, Bacterial - chemistry
Exact sciences and technology
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Genomics
Humans
In vitro gene amplification. Pcr technique
Mass spectrometry
Mass Spectrometry - methods
Methods. Procedures. Technologies
Other chromatographic methods
Plasma - microbiology
Polymerase Chain Reaction - methods
Reproducibility of Results
Sensitivity and Specificity
Spectrometric and optical methods
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Summary:Bloodstream infections are an important cause of serious morbidity and mortality. Rapid detection and identification of specific pathogens from blood or other clinical specimens could improve the rational use of antimicrobial therapy in clinical medicine and have a great impact on the outcome of patients with systemic infections. Polymerase chain reaction using generic primers was used to amplify genomic DNA of different bacterial strains. The identification was accomplished by measuring the molecular masses of the PCR products using ion-pair reversed-phase high-performance liquid chromatography hyphenated to electrospray ionization mass spectrometry. DNA from 10 bacterial species was amplified by PCR, and the resulting amplification products were analyzed. In all cases, the measured molecular masses of the PCR products matched the theoretical value for the species-specific DNA sequence. However, three pairs of bacteria could not be distinguished since the theoretical difference in amplicon molecular mass was
ISSN:0003-2700
1520-6882
DOI:10.1021/ac050378l