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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

  • Environmental Microbiology - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

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.

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Acknowledgements

We thank Drs. Huizhong Chen and Kidon Sung for critical review of the manuscript. The views expressed herein do not necessarily reflect those of the U.S. Food and Drug Administration or the U.S. Department of Health and Human Services.

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Authors and Affiliations

  1. Division of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, 3900 NCTR Road, Jefferson, AR, 72079-9502, USA

    Youngbeom Ahn, Bailey Gibson & Carl E. Cerniglia

  2. Division of Systems Biology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR, 72079, USA

    Anna Williams, Pierre Alusta & Dan A. Buzatu

  3. Department of Biological Sciences, Albany State University, Albany, GA, 31705, USA

    Yong-Jin Lee

  4. Department of Pediatrics, University of Michigan, Ann Arbor, MI, 48109, USA

    John J. LiPuma

  5. Eagle Analytical Services, Houston, TX, 77099, USA

    David Hussong

  6. Office of Pharmaceutical Quality, Center for Drug Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, MD, 20993, USA

    Bernard Marasa

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  1. Youngbeom Ahn
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Ahn, Y., Gibson, B., Williams, A. et al. 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 47, 475–484 (2020). https://doi.org/10.1007/s10295-020-02287-3

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  • DOI: https://doi.org/10.1007/s10295-020-02287-3

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