Comparison of three rapid and easy bacterial DNA extraction methods for use with quantitative real-time PCR

S. P. Tongeren (1) , J. E. Degener (1) , H. J. M. Harmsen (1)

1 Department of Medical Microbiology

The development of fast and easy on-site molecular detection and quantification methods for hazardous microbes on solid surfaces is desirable for several applications where specialised laboratory facilities are absent. The quantification of bacterial contamination necessitates the assessment of the efficiency of the used methodology as a whole, including the preceding steps of sampling and sample processing. We used quantitative real-time polymerase chain reaction (qrtPCR) for and to measure the recovery of DNA from defined numbers of bacterial cells that were subjected to three different DNA extraction methods: the QIAamp® DNA Mini Kit, Reischl et al.'s method and FTA® Elute. FTA® Elute significantly showed the highest median DNA extraction efficiency of 76.9% for and 108.9% for . The Reischl et al. method and QIAamp® DNA Mini Kit inhibited the qrtPCR assay with a 10-fold decrease of detectable DNA. None of the methods inhibited the qrtPCR assay. The FTA® Elute applicability was demonstrated with swab samples taken from the International Space Station (ISS) interior. Overall, the FTA® Elute method was found to be the most suitable to selected criteria in terms of rapidity, easiness of use, DNA extraction efficiency, toxicity, and transport and storage conditions.

Mots clés: Life Sciences

S. P. Tongeren, J. E. Degener, H. J. M. Harmsen. Comparison of three rapid and easy bacterial DNA extraction methods for use with quantitative real-time PCR. European Journal of Clinical Microbiology and Infectious Diseases, 2011, 30 (9), pp.1053-1061. ⟨10.1007/s10096-011-1191-4⟩. ⟨hal-00669043⟩