Control of the global epidemic tuberculosis is severely hampered by the emergence of drug-resistant Mycobacterium tuberculosis strains. Molecular methods offer a more rapid means of characterizing resistant strains than phenotypic drug susceptibility testing. We have developed a molecular method for detection of rifampicin-resistant M. tuberculosis based on padlock probes and magnetic nanobeads. Padlockprobes were designed to target the most common mutations associated with rifampicinresistance in M. tuberculosis, i.e. at codons 516, 526 and 531 in the gene rpoB. Fordetection of the wild type sequence at all three codons simultaneously, a padlock probe and two gap-fill oligonucleotides were used in a novel assay configuration, requiring three ligation events for circularization. The assay also includes a probe for identificationof the M. tuberculosis complex. Circularized probes were amplified by rolling circle amplification. Amplification products were coupled to oligonucleotide-conjugatedmagnetic nanobeads and detected by measuring the frequency-dependent magneticresponse of the beads using a portable AC susceptometer.
| Identifer | oai:union.ndltd.org:UPSALLA1/oai:DiVA.org:uu-198729 |
| Date | January 2013 |
| Creators | Engström, Anna, Zardán Gómez de la Torre, Teresa, Strømme, Maria, Nilsson, Mats, Herthnek, David |
| Publisher | Uppsala universitet, Nanoteknologi och funktionella material, Uppsala universitet, Nanoteknologi och funktionella material, Uppsala universitet, Science for Life Laboratory, SciLifeLab, Uppsala universitet, Science for Life Laboratory, SciLifeLab, Uppsala universitet, Molekylära verktyg |
| Source Sets | DiVA Archive at Upsalla University |
| Language | English |
| Detected Language | English |
| Type | Article in journal, info:eu-repo/semantics/article, text |
| Format | application/pdf |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | PLoS ONE, 1932-6203, 2013, 8:4, s. e62015- |
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