Identification and typing of Francisella tularensis with a highly automated genotyping assay

D Duncan, A Vogler, M Wolcott, F Li, Derek Sarovich, Dawn Birdsell, Lindsey Watson, T Hall, Rangarajan Sampath, R Housley, L Blyn, S Hofstadler, David Ecker, Paul S Keim, David M Wagner, Mark Eshoo

    Research output: Contribution to journalArticle

    Abstract

    A PCR assay was developed to genotypically characterize Francisella tularensis and F. novicida. An integrated and partially redundant set of markers was selected to provide positive identification of these species, identify subspecies of F. tularensis and genotype 14 variable number tandem repeat (VNTR) markers. Assay performance was evaluated with 117 Francisella samples. Sample DNA was amplified, and the masses of the PCR products were determined with electrospray ionization/time of flight mass spectrometry (ESI-MS). The base compositions of the PCR amplicons were derived from these high-accuracy mass measurements and contrasted with databased information associated with each of the 25 assay markers. Species and subspecies determinations for all samples were fully concordant with results from established typing methods, and VNTR markers provided additional discrimination among samples. Sequence variants were observed with a number of assay markers, but these did not interfere with sample characterization, and served to increase the genetic diversity detected by the assay.
    Original languageEnglish
    Pages (from-to)128-134
    Number of pages7
    JournalLetters in Applied Microbiology
    Volume56
    Issue number2
    DOIs
    Publication statusPublished - 2013

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    Duncan, D., Vogler, A., Wolcott, M., Li, F., Sarovich, D., Birdsell, D., Watson, L., Hall, T., Sampath, R., Housley, R., Blyn, L., Hofstadler, S., Ecker, D., Keim, P. S., Wagner, D. M., & Eshoo, M. (2013). Identification and typing of Francisella tularensis with a highly automated genotyping assay. Letters in Applied Microbiology, 56(2), 128-134. https://doi.org/10.1111/lam.12022