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DNA Extraction from Clinical Specimens for the Direct Detection of Mycobacterium ulcerans by Real-Time PCR
Caroline J Lavender1, Janet A M Fyfe2
1Victorian Infectious Diseases Reference Laboratory, Doherty Institute, Melbourne, Victoria, Australia.
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 .
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