Proposition of a safe Mycobacterium tuberculosis complex denaturation method that does not compromise the integrity of DNA for whole-genome sequencing.
Bacteriological Techniques
Chloroform
/ pharmacology
DNA, Bacterial
/ analysis
Ethanol
/ pharmacology
Humans
Laboratory Infection
/ prevention & control
Microbial Viability
/ drug effects
Mycobacterium tuberculosis
/ drug effects
Safety Management
/ methods
Specimen Handling
/ methods
Tuberculosis
/ microbiology
Whole Genome Sequencing
/ methods
Denaturation
Mycobacterium tuberculosis
Whole-genome sequencing
Journal
Tuberculosis (Edinburgh, Scotland)
ISSN: 1873-281X
Titre abrégé: Tuberculosis (Edinb)
Pays: Scotland
ID NLM: 100971555
Informations de publication
Date de publication:
07 2019
07 2019
Historique:
received:
12
12
2018
revised:
18
06
2019
accepted:
19
06
2019
entrez:
6
8
2019
pubmed:
6
8
2019
medline:
18
2
2020
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 31378270
pii: S1472-9792(18)30518-3
doi: 10.1016/j.tube.2019.06.003
pii:
doi:
Substances chimiques
DNA, Bacterial
0
Ethanol
3K9958V90M
Chloroform
7V31YC746X
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
62-64Informations de copyright
Copyright © 2019 Elsevier Ltd. All rights reserved.