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Prospective evaluation of pyrosequencing for the rapid detection of isoniazid and rifampin resistance in clinical Mycobacterium tuberculosis isolates

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Abstract

A pyrosequencing-based method for the rapid detection of isoniazid (INH) and rifampin (RIF) resistance in Mycobacterium tuberculosis was evaluated in clinical practice. The method can detect the INH resistance-causing katG315 mutation, and all mutations in the RIF resistance-determining rpoB core region, in less than 6 h from cultured isolates. The method was first validated with 42 isolates, and was subsequently prospectively evaluated with 91 isolates, including clinical isolates and external quality control assessment strains, over a period of 2.5 years. The pyrosequencing results of clinical isolates were available, on average, 19 days earlier (median 19 days; range 3–43 days) than conventional susceptibility testing results. The composite data showed that the sensitivity of pyrosequencing for detecting resistance correctly was 66.7% for INH and 97.4% for RIF. The specificity of pyrosequencing was 100% for both drugs. Acceptable sensitivity for detecting resistance and the rapidness of pyrosequencing make it a valuable tool in the clinical setting.

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Acknowledgments

We thank Marita Kirjonen, Eija Lönnblad, Anna Musku, Tiina Nurmio, and Ulla Toivonen for their excellent technical assistance.

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  1. Mycobacterial Reference Laboratory, National Public Health Institute, Kiinamyllynkatu 13, 20520, Turku, Finland

    H. J. Marttila, J. Mäkinen, M. Marjamäki & H. Soini

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  1. H. J. Marttila
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Correspondence to J. Mäkinen.

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Marttila, H.J., Mäkinen, J., Marjamäki, M. et al. Prospective evaluation of pyrosequencing for the rapid detection of isoniazid and rifampin resistance in clinical Mycobacterium tuberculosis isolates. Eur J Clin Microbiol Infect Dis 28, 33–38 (2009). https://doi.org/10.1007/s10096-008-0584-5

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  • DOI: https://doi.org/10.1007/s10096-008-0584-5

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