Background: Pathogen identification in clinical routine is based on the cultivation of microbes with subsequent morphological and physiological characterisation lasting at least 24 hours. However, early and accurate identification is a crucial requisite for fast and optimally targeted antimicrobial treatment. Molecular biology based techniques allow fast identification, however discrimination of very closely related species remains still difficult. Results: A molecular approach is presented for the rapid identification of pathogens combining PCR amplification with microarray detection. The DNA chip comprises oligonucleotide capture probes for 25 different pathogens including Gram positive cocci, the most frequently encountered genera of Enterobacteriaceae, non- fermenter and clinical relevant Candida species. The observed detection limits varied from 10 cells ( e. g. E. coli) to 105 cells ( S. aureus) per mL artificially spiked blood. Thus the current low sensitivity for some species still represents a barrier for clinical application. Successful discrimination of closely related species was achieved by a signal pattern recognition approach based on the k- nearest- neighbour method. A prototype software providing this statistical evaluation was developed, allowing correct identification in 100 % of the cases at the genus and in 96.7 % at the species level ( n = 241). Conclusion: The newly developed molecular assay can be carried out within 6 hours in a research laboratory from pathogen isolation to species identification. From our results we conclude that DNA microarrays can be a useful tool for rapid identification of closely related pathogens particularly when the protocols are adapted to the special clinical scenarios.
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Belgrader, P
;
Benett, W
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Benett, W
;
Hadley, D
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Hadley, D
;
Richards, J
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Richards, J
;
Stratton, P
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Stratton, P
;
Mariella, R
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Mariella, R
;
Milanovich, F
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
机构:
ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, AustriaARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Bodrossy, L
;
Stralis-Pavese, N
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Stralis-Pavese, N
;
Murrell, JC
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Murrell, JC
;
Radajewski, S
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Radajewski, S
;
Weilharter, A
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Weilharter, A
;
Sessitsch, A
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
机构:
Guys & St Thomas Hosp Trust, St Thomas Hosp, Dept Microbiol, London SE1 7EH, EnglandGuys & St Thomas Hosp Trust, St Thomas Hosp, Dept Microbiol, London SE1 7EH, England
Brown, TJ
;
Anthony, RM
论文数: 0引用数: 0
h-index: 0
机构:Guys & St Thomas Hosp Trust, St Thomas Hosp, Dept Microbiol, London SE1 7EH, England
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Belgrader, P
;
Benett, W
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Benett, W
;
Hadley, D
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Hadley, D
;
Richards, J
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Richards, J
;
Stratton, P
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Stratton, P
;
Mariella, R
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
Mariella, R
;
Milanovich, F
论文数: 0引用数: 0
h-index: 0
机构:
Univ Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USAUniv Calif Lawrence Livermore Natl Lab, Biol & Biotechnol Res Program, Livermore, CA 94551 USA
机构:
ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, AustriaARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Bodrossy, L
;
Stralis-Pavese, N
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Stralis-Pavese, N
;
Murrell, JC
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Murrell, JC
;
Radajewski, S
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Radajewski, S
;
Weilharter, A
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
Weilharter, A
;
Sessitsch, A
论文数: 0引用数: 0
h-index: 0
机构:ARC Seibersdorf Res GmbH, Dept Biotechnol, Div Life & Environm Sci, A-2444 Seibersdorf, Austria
机构:
Guys & St Thomas Hosp Trust, St Thomas Hosp, Dept Microbiol, London SE1 7EH, EnglandGuys & St Thomas Hosp Trust, St Thomas Hosp, Dept Microbiol, London SE1 7EH, England
Brown, TJ
;
Anthony, RM
论文数: 0引用数: 0
h-index: 0
机构:Guys & St Thomas Hosp Trust, St Thomas Hosp, Dept Microbiol, London SE1 7EH, England