Proposition of a safe Mycobacterium tuberculosis complex denaturation method that does not compromise the integrity of DNA for whole-genome sequencing

Tuberculosis
Typhaine Billard-PomaresEtienne Carbonnelle

Abstract

Whole-genome sequencing plays now a leading role in epidemiologic studies of tuberculosis. DNA extraction of Mycobacterium tuberculosis complex (MTBC) requires complete inactivation of the strains, to be handled for further molecular procedures. In this study we compared two chloroform-based denaturation methods (one with a step of heat killing, one without) to a traditional heat inactivation method. Our results showed that 40% of the strains of MTBC treated by the traditional protocol resulted in a positive culture whereas no culture was observed with the two chloroform-based protocols. The DNA extracts obtained with chloroform-based protocols preparation were successfully used for whole-genome sequencing. We recommend inactivation with our rapid and efficient denaturation method using chloroform without heat killing which met our expectations and biosecurity requirements.

References

Jun 12, 1999·Journal of Clinical Microbiology·P Bemer-Melchior, H B Drugeon
Oct 2, 2002·Journal of Clinical Pathology·C DoigK J Forbes
Jun 16, 2005·Journal of Clinical Microbiology·Wendy SomervilleMarcel A Behr
Mar 10, 2017·Journal of Clinical Microbiology·Antonina A VotintsevaZamin Iqbal
Mar 17, 2017·Journal of Clinical Microbiology·Robyn S Lee, Madhukar Pai

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Citations

Jul 29, 2020·Journal of Clinical Microbiology·Sophie GeorgeKate E Dingle

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