A novel, multiplex, real-time PCR-based approach for the detection of the commonly occurring pathogenic fungi and bacteria

BMC Microbiology
Ádám HorváthFerenc Somogyvári

Abstract

Polymerase chain reaction (PCR)-based techniques are widely used to identify fungal and bacterial infections. There have been numerous reports of different, new, real-time PCR-based pathogen identification methods although the clinical practicability of such techniques is not yet fully clarified.The present study focuses on a novel, multiplex, real-time PCR-based pathogen identification system developed for rapid differentiation of the commonly occurring bacterial and fungal causative pathogens of bloodstream infections. A multiplex, real-time PCR approach is introduced for the detection and differentiation of fungi, Gram-positive (G+) and Gram-negative (G-) bacteria. The Gram classification is performed with the specific fluorescence resonance energy transfer (FRET) probes recommended for LightCycler capillary real-time PCR. The novelty of our system is the use of a non-specific SYBR Green dye instead of labelled anchor probes or primers, to excite the acceptor dyes on the FRET probes. In conjunction with this, the use of an intercalating dye allows the detection of fungal amplicons.With the novel pathogen detection system, fungi, G + and G- bacteria in the same reaction tube can be differentiated within an hour after the DNA ...Continue Reading

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Citations

Nov 19, 2015·Expert Review of Molecular Diagnostics·Robert DuncanMing Dong
Nov 9, 2016·MSystems·Maxime GalanJean-François Cosson
Jan 16, 2020·PLoS Neglected Tropical Diseases·Amir ArastehfarSarah A Ahmed
Oct 13, 2019·Microbial Biotechnology·Yannick WoutersGeert J A Wanten
Jun 15, 2019·Laboratory Medicine·Nada N NawarMohamed A Emara
Feb 18, 2021·FEMS Microbiology Reviews·Wouter Van GenechtenLiesbeth Demuyser
Dec 12, 2019·ACS Applied Materials & Interfaces·Haitao YuanShu Wang

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Methods Mentioned

BETA
PCR
fluorescence resonance
FRET
electrophoresis

Software Mentioned

LightCycler

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