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Fluoroquinolone heteroresistance in Mycobacterium tuberculosis: detection by genotypic and phenotypic assays in experimentally mixed populations

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dc.contributor.author Rigouts, L. en_US
dc.contributor.author Miotto, P. en_US
dc.contributor.author Schats, M. en_US
dc.contributor.author Lempens, P. en_US
dc.contributor.author Cabibbe, A. M. en_US
dc.contributor.author Galbiati, S. en_US
dc.contributor.author Lampasona, V. en_US
dc.contributor.author De Rijk, P. en_US
dc.contributor.author Cirillo, D. M. en_US
dc.contributor.author de Jong, B. C. en_US
dc.date.accessioned 2020-08-25T09:38:42Z
dc.date.available 2020-08-25T09:38:42Z
dc.date.issued 2019 en_US
dc.identifier.issn 2045-2322 en_US
dc.identifier.doi http://dx.doi.org/10.1038/s41598-019-48289-9 en_US
dc.identifier.other http://lib.itg.be/pdf/itg/2019/2019srep0002.pdf en_US
dc.identifier.other 8 pp. en_US
dc.identifier.other 33 en_US
dc.identifier.other ITG-B1A; ITG-B8A; ITG-BLA; DBM; U-MYCOB; JIF; DOI; PDF; PMC; Abstract; ITMPUB; DSPACE68 en_US
dc.identifier.uri http://hdl.handle.net/10390/10946
dc.description.abstract Heteroresistance - the simultaneous presence of drug-susceptible and -resistant organisms - is common in Mycobacterium tuberculosis. In this study, we aimed to determine the limit of detection (LOD) of genotypic assays to detect gatifloxacin-resistant mutants in experimentally mixed populations. A fluoroquinolone-susceptible M. tuberculosis mother strain (S) and its in vitro selected resistant daughter strain harbouring the D94G mutation in gyrA (R) were mixed at different ratio's. Minimum inhibitory concentrations (MICs) against gatifloxacin were determined, while PCR-based techniques included: line probe assays (Genotype MTBDRsl and GenoScholar-FQ + KM TB II), Sanger sequencing and targeted deep sequencing. Droplet digital PCR was used as molecular reference method. A breakpoint concentration of 0.25 mg/L allows the phenotypic detection of >/=1% resistant bacilli, whereas at 0.5 mg/L >/= 5% resistant bacilli are detected. Line probe assays detected >/=5% mutants. Sanger sequencing required the presence of around 15% mutant bacilli to be detected as (hetero) resistant, while targeted deep sequencing detected </=1% mutants. Deep sequencing and phenotypic testing are the most sensitive methods for detection of fluoroquinolone-resistant minority populations, followed by line probe assays (provided that the mutation is confirmed by a mutation band), while Sanger sequencing proved to be the least sensitive method. en_US
dc.language English en_US
dc.relation.uri http://www.ncbi.nlm.nih.gov/pubmed/31409849 en_US
dc.subject Mycobacterium tuberculosis en_US
dc.subject Bacteriology en_US
dc.subject Drug resistance en_US
dc.title Fluoroquinolone heteroresistance in Mycobacterium tuberculosis: detection by genotypic and phenotypic assays in experimentally mixed populations en_US
dc.type Article-E en_US
dc.citation.issue 1 en_US
dc.citation.jtitle Scientific Reports en_US
dc.citation.volume 9 en_US
dc.citation.pages 11760 en_US
dc.citation.abbreviation Sci Rep en_US


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