Broad-range PCR for the identification of bacterial and fungal pathogens from blood: a sequencing approach

Methods Mol Biol. 2015:1237:129-38. doi: 10.1007/978-1-4939-1776-1_13.

Abstract

Broad-range PCR has become a valuable tool for the identification of microorganisms in the clinical laboratory over the last years. It was primarily used to identify slow-growing and fastidious microorganisms with poor biochemical activity. Nowadays, it is also used to identify microorganisms directly from clinical samples such as blood or punctuates from primarily sterile body sites. In these specimens, the usage of broad-range PCR is challenging regarding contamination and standardization. To overcome these problems, a new test system, the SepsiTest™, was introduced recently employing broad-range PCR for the identification of microorganisms in septic patients. In this chapter, the test system is described and the equipment necessary listed.

MeSH terms

  • Automation, Laboratory
  • Bacteremia / diagnosis*
  • Bacteremia / microbiology
  • Bacteremia / pathology
  • Bacteria / genetics
  • Bacteria / growth & development
  • Bacteria / isolation & purification
  • Blood Specimen Collection
  • Culture Media / chemistry
  • DNA, Bacterial / genetics
  • DNA, Bacterial / isolation & purification*
  • DNA, Fungal / genetics
  • DNA, Fungal / isolation & purification*
  • Fungemia / diagnosis*
  • Fungemia / microbiology
  • Fungemia / pathology
  • Fungi / genetics
  • Fungi / growth & development
  • Fungi / isolation & purification
  • High-Throughput Nucleotide Sequencing
  • Humans
  • Molecular Diagnostic Techniques / instrumentation
  • Molecular Diagnostic Techniques / methods*
  • Polymerase Chain Reaction / instrumentation
  • Polymerase Chain Reaction / methods*
  • Sequence Analysis, DNA

Substances

  • Culture Media
  • DNA, Bacterial
  • DNA, Fungal