Abstract
For patients infected with tuberculosis, detection of rpoB gene mutations is critical for diagnosing drug-resistant strains of the causative pathogen, Mycobacterium tuberculosis (MTB). Traditional approaches to drug resistance include culture, which is very slow. Recently described real-time polymerase chain reaction approaches have addressed turnaround time but at relatively high cost. Here, we describe a novel amplification method, termed blocked-primer helicase-dependent amplification, for amplifying rpoB gene sequences in MTB. Resultant amplicon is hybridized to a probe set arrayed on a modified silicon-based chip to determine if there is any mutation in that region. Using this method, we could detect the majority of clinically relevant mutations in rpoB gene.
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Acknowledgments
We thank Adrianne Clifford and all other members of the Research Group at Great Basin Corporation for technical assistance and Steve Aldous for financial support for this project.
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Ao, W., Jenison, R. (2013). Detection of rpoB Gene Mutations Using Helicase-Dependent Amplification. In: Kolpashchikov, D., Gerasimova, Y. (eds) Nucleic Acid Detection. Methods in Molecular Biology, vol 1039. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-535-4_7
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DOI: https://doi.org/10.1007/978-1-62703-535-4_7
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