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Molecular beacon sequence analysis for detecting drug resistance in Mycobacterium tuberculosis

Nature Biotechnology volume 16, pages 359–363 (1998)Cite this article

Abstract

We developed a new approach to DNA sequence analysis that uses fluorogenic reporter molecules—molecular beacons—and demonstrated their ability to discriminate alleles in real-time PCR assays of genomic DNA. A set of overlapping molecular beacons was used to analyze an 81-bp region of the Mycobacterium tuberculosis rpoB gene for mutations that confer resistance to the antibiotic rifampin. In a blinded study of 52 rifampin-resistant and 23 rifampin-susceptible clinical isolates, this method correctly detected mutations in all of the resistant strains and in none of the susceptible strains. The assay was carried out entirely in sealed PCR tubes and was simple to perform and interpret. This approach can be used to analyze any DNA sequence of moderate length with single base pair accuracy.

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Authors and Affiliations

  1. Division of Infectious Diseases, Department of Medicine, Montefiore Medical Center, Bronx, NY, 10467

    Amy S. Piatek & David Alland

  2. Department of Molecular Genetics, Public Health Research Institute, New York, NY, 10016

    Sanjay Tyagi & Fred Russell Kramer

  3. Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY, 10461

    Arno C. Pol

  4. Division of Infectious Diseases, Centre Hospitalier Universitaire Vaudois, 1011, Lausanne, Switzerland

    Amalio Telenti

  5. Department of Medicine, University of Medicine and Dentistry of New Jersey, New Jersey Medical School, Newark, NJ, 07103

    Lincoln P. Miller

Authors
  1. Amy S. Piatek
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  2. Sanjay Tyagi
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  3. Arno C. Pol
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  4. Amalio Telenti
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  5. Lincoln P. Miller
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  6. Fred Russell Kramer
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  7. David Alland
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Correspondence to David Alland.

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Piatek, A., Tyagi, S., Pol, A. et al. Molecular beacon sequence analysis for detecting drug resistance in Mycobacterium tuberculosis. Nat Biotechnol 16, 359–363 (1998). https://doi.org/10.1038/nbt0498-359

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