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Molecular drug susceptibility testing and strain typing of tuberculosis by DNA hybridization.


ABSTRACT: In Mycobacterium tuberculosis (Mtb) the detection of single nucleotide polymorphisms (SNPs) is of high importance both for diagnostics, since drug resistance is primarily caused by the acquisition of SNPs in multiple drug targets, and for epidemiological studies in which strain typing is performed by SNP identification. To provide the necessary coverage of clinically relevant resistance profiles and strain types, nucleic acid-based measurement techniques must be able to detect a large number of potential SNPs. Since the Mtb problem is pressing in many resource-poor countries, requiring low-cost point-of-care biosensors, this is a non-trivial technological challenge. This paper presents a proof-of-concept in which we chose simple DNA-DNA hybridization as a sensing principle since this can be transferred to existing low-cost hardware platforms, and we pushed the multiplex boundaries of it. With a custom designed probe set and a physicochemical-driven data analysis it was possible to simultaneously detect the presence of SNPs associated with first- and second-line drug resistance and Mtb strain typing. We have demonstrated its use for the identification of drug resistance and strain type from a panel of phylogenetically diverse clinical strains. Furthermore, reliable detection of the presence of a minority population (<5%) of drug-resistant Mtb was possible.

SUBMITTER: Wood HN 

PROVIDER: S-EPMC6366778 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Molecular drug susceptibility testing and strain typing of tuberculosis by DNA hybridization.

Wood Hillary N HN   Venken Tom T   Willems Hanny H   Jacobs An A   Reis Ana Júlia AJ   Almeida da Silva Pedro Eduardo PE   Homolka Susanne S   Niemann Stefan S   Rohde Kyle H KH   Hooyberghs Jef J  

PloS one 20190207 2


In Mycobacterium tuberculosis (Mtb) the detection of single nucleotide polymorphisms (SNPs) is of high importance both for diagnostics, since drug resistance is primarily caused by the acquisition of SNPs in multiple drug targets, and for epidemiological studies in which strain typing is performed by SNP identification. To provide the necessary coverage of clinically relevant resistance profiles and strain types, nucleic acid-based measurement techniques must be able to detect a large number of  ...[more]

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