Fungal-Grade Reagents and Materials for Molecular Analysis

Methods Mol Biol. 2017:1508:141-150. doi: 10.1007/978-1-4939-6515-1_6.

Abstract

Fungal DNA is present at very low loads in clinical specimens. Molecular detection by amplification assays generally is a challenge because of a potentially multiple input of contaminating DNA from exogenous sources. Besides airborne, handling and cross-contamination, materials and reagents used in the molecular laboratory can contain microbial DNA which is a long underestimated potential source of false positive results. In this contribution decontamination procedures of materials and reagents and the selection of certified microbial DNA-free components for sample collection, DNA extraction, and PCR amplification are discussed with respect to the aim of building up a reliable molecular system for the diagnosis of fungal organisms at the limit of detection.

Keywords: Contamination; DNA-free; Decontamination; Fungal DNA; Fungal DNA extraction; Fungal PCR reagents; Fungal assays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • DNA Contamination
  • DNA, Fungal / genetics
  • DNA, Fungal / isolation & purification*
  • Fungi / genetics*
  • Humans
  • Molecular Diagnostic Techniques*
  • Mycoses / diagnosis*
  • Mycoses / microbiology

Substances

  • DNA, Fungal