Skip to main content

DNA Extraction from Clinical Specimens for the Direct Detection of Mycobacterium ulcerans by Real-Time PCR

  • Protocol
  • First Online:
Mycobacterium ulcerans

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2387))

Abstract

Mycobacterium ulcerans is a slow-growing environmental bacterium that causes a severe skin disease known as Buruli ulcer (BU). Rapid detection of M. ulcerans in clinical specimens is essential for early diagnosis so that patients can be treated appropriately as soon as possible. This chapter describes suitable methods for the extraction of M. ulcerans DNA from the most common specimens submitted to the laboratory for confirmation of BU: swabs, fresh tissue biopsies, and fixed tissue sections. The resulting DNA extracts may be used for downstream procedures including standard gel-based PCR and real-time PCR assays. Protocols for direct detection of M. ulcerans DNA by real-time PCR are described in Chapter 8.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. MacCallum P, Tolhurst J, Buckle G, Sissons H (1948) A new mycobacterial infection in man. J Path Bacteriol 60:93–122

    Article  CAS  Google Scholar 

  2. Fyfe JA, Lavender CJ, Johnson PD et al (2007) Development and application of two multiplex real-time PCR assays for the detection of Mycobacterium ulcerans in clinical and environmental samples. Appl Environ Microbiol 73:4733–4740

    Article  CAS  Google Scholar 

  3. Phillips R, Horsfield C, Kuijper S et al (2005) Sensitivity of PCR targeting the IS2404 insertion sequence of Mycobacterium ulcerans in an assay using punch biopsy specimens for diagnosis of Buruli ulcer. J Clin Microbiol 43:3650–3656

    Article  CAS  Google Scholar 

  4. Ross B, Marino L, Oppedisano F et al (1997) Development of a PCR assay for rapid diagnosis of Mycobacterium ulcerans infection. J Clin Microbiol 35:1696–1700

    Article  CAS  Google Scholar 

  5. Portaels F, Johnson P, Meyers WM (2001) Buruli ulcer; diagnosis of Mycobacterium ulcerans disease; a manual for health care providers. World Health Organization, Geneva. http://www.who.int/buruli/information/diagnosis/en/index.html

    Google Scholar 

  6. Portaels F, Meyers WM, Ablordey A et al (2008) First cultivation and characterization of Mycobacterium ulcerans from the environment. PLoS Negl Trop Dis 2:e178

    Article  Google Scholar 

  7. Durnez L, Stragier P, Roebben K et al (2008) A comparison of DNA extraction procedures for the detection of Mycobacterium ulcerans, the causative agent of Buruli ulcer, in clinical and environmental specimens. J Microbiol Methods 76:152–158

    Article  Google Scholar 

  8. Affolabi D, Sanoussi N, Vandelannoote K et al (2012) Effects of decontamination, DNA extraction, and amplification procedures on the molecular diagnosis of Mycobacterium ulcerans disease (Buruli Ulcer). J Clin Microbiol 50:1195–1198

    Article  CAS  Google Scholar 

  9. Radomski N, Kreitmann L, McIntosh F, Beyr M (2013) The critical role of DNA extraction of mycobacteria in tissues. PLoS One 8:e78749

    Article  CAS  Google Scholar 

  10. Eddyani M, Lavender C, de Rijk WB, Bomans P, Fyfe J, de Jong B et al (2014) Multicenter external quality assessment program for PCR detection of Mycobacterium ulcerans in clinical and environmental specimens. PLoS One 9(2):e89407

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Janet A. M. Fyfe .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Lavender, C.J., Fyfe, J.A.M. (2022). DNA Extraction from Clinical Specimens for the Direct Detection of Mycobacterium ulcerans by Real-Time PCR. In: Pluschke, G., Röltgen, K. (eds) Mycobacterium ulcerans. Methods in Molecular Biology, vol 2387. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1779-3_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1779-3_6

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1778-6

  • Online ISBN: 978-1-0716-1779-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics