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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
MacCallum P, Tolhurst J, Buckle G, Sissons H (1948) A new mycobacterial infection in man. J Path Bacteriol 60:93–122
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
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
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
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
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
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
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
Radomski N, Kreitmann L, McIntosh F, Beyr M (2013) The critical role of DNA extraction of mycobacteria in tissues. PLoS One 8:e78749
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
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
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