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Sepsis pp 57-64 | Cite as

Bacterial and Fungal DNA Extraction from Positive Blood Culture Bottles: A Manual and an Automated Protocol

  • Minna MäkiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1237)

Abstract

When adapting a gene amplification-based method in a routine sepsis diagnostics using a blood culture sample as a specimen type, a prerequisite for a successful and sensitive downstream analysis is the efficient DNA extraction step. In recent years, a number of in-house and commercial DNA extraction solutions have become available. Careful evaluation in respect to cell wall disruption of various microbes and subsequent recovery of microbial DNA without putative gene amplification inhibitors should be conducted prior selecting the most feasible DNA extraction solution for the downstream analysis used. Since gene amplification technologies have been developed to be highly sensitive for a broad range of microbial species, it is also important to confirm that the used sample preparation reagents and materials are bioburden-free to avoid any risks for false-positive result reporting or interference of the diagnostic process. Here, one manual and one automated DNA extraction system feasible for blood culture samples are described.

Key words

Sample preparation Blood culture DNA extraction Fungi Bacteria 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Orion Diagnostica OyEspooFinland

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