A comparison of culture-based, real-time PCR, droplet digital PCR and flow cytometric methods for the detection of Burkholderia cepacia complex in nuclease-free water and antiseptics

J Ind Microbiol Biotechnol. 2020 Jul;47(6-7):475-484. doi: 10.1007/s10295-020-02287-3. Epub 2020 Jul 15.

Abstract

The presence of Burkholderia cepacia complex (BCC) strains has resulted in recalls of pharmaceutical products, since these opportunistic pathogens can cause serious infections. Rapid and sensitive diagnostic methods to detect BCC are crucial to determine contamination levels. We evaluated bacterial cultures, real-time PCR (qPCR), droplet digital PCR (ddPCR), and flow cytometry to detect BCC in nuclease-free water, in chlorhexidine gluconate (CHX) and benzalkonium chloride (BZK) solutions. Twenty BCC strains were each suspended (1, 10, 100, and 1000 CFU/ml) in autoclaved nuclease-free water, 10 μg/ml CHX, and 50 μg/ml BZK. Five replicates of each strain were tested at each concentration (20 strains × 4 concentrations × 5 replicates = 400 tests) to detect BCC using the aforementioned four methods. We demonstrated the potential of ddPCR and flow cytometry as more sensitive alternatives to culture-based methods to detect BCC in autoclaved nuclease-free water and antiseptics samples.

Keywords: Antiseptics; Burkholderia cepacia complex; Droplet digital PCR; Flow cytometers; Nuclease-free water; Real-time PCR.

Publication types

  • Comparative Study

MeSH terms

  • Anti-Infective Agents, Local / pharmacology*
  • Benzalkonium Compounds
  • Biotechnology
  • Burkholderia cepacia complex*
  • Chlorhexidine / analogs & derivatives
  • Culture
  • Drug Contamination*
  • Flow Cytometry*
  • Polymerase Chain Reaction / methods*
  • Real-Time Polymerase Chain Reaction*
  • Water

Substances

  • Anti-Infective Agents, Local
  • Benzalkonium Compounds
  • Water
  • chlorhexidine gluconate
  • Chlorhexidine