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High-throughput screening of bacterial pathogens in clinical specimens using 16S rDNA qPCR and fragment analysis.
Journal article

High-throughput screening of bacterial pathogens in clinical specimens using 16S rDNA qPCR and fragment analysis.

  • Wagner K Institute of Medical Microbiology, University of Zurich, Zurich, Switzerland. Electronic address: karoline.wagner@usb.ch.
  • Springer B Institute of Medical Microbiology and Hygiene, Austrian Agency for Health and Food Safety, Graz, Austria.
  • Pires VP Institute of Medical Microbiology, University of Zurich, Zurich, Switzerland.
  • Keller PM Institute of Medical Microbiology, University of Zurich, Zurich, Switzerland.
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  • 2018-12-15
Published in:
  • Diagnostic microbiology and infectious disease. - 2019
16S
Bacterial broad-range qPCR
Bacterial infection
Fragment analysis
High throughput screening
Bacteria
Bacterial Infections
Bacteriological Techniques
Humans
Mass Screening
Molecular Diagnostic Techniques
RNA, Ribosomal, 16S
Real-Time Polymerase Chain Reaction
Retrospective Studies
Switzerland
English Molecular-based detection of bacterial pathogens directly from clinical specimens permits rapid initiation of effective antimicrobial treatment and adequate patient management. Broad-range polymerase chain reaction (PCR) amplification of the 16S rRNA gene (16S rDNA qPCR) is used in many diagnostic laboratories as a complement to cultural identification of bacterial pathogens. However, efforts for automation of 16S rDNA PCR workflows are needed in order to reduce turnaround times and to enhance reproducibility and standardization of the technique. In this retrospective method evaluation study, clinical specimens (N = 499) from patients with suspected bacterial infections were used to evaluate 2 diagnostic semiautomated workflows for rapid bacterial pathogen detection. The workflows included automated DNA extraction (QIASymphony), 16S rDNA qPCR, fragment or melting curve analysis, and amplicon sequencing. Our results support the use of the 16S rDNA qPCR and fragment analysis workflow as it enabled rapid and accurate identification of bacterial pathogens in clinical specimens.
Language
  • English
Open access status
hybrid
Identifiers
Persistent URL
https://sonar.ch/global/documents/271128
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