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Characterization of Clostridium species utilizing liquid chromatography/mass spectrometry of intact proteins.

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      Abstract

      A method for biomarker candidate discovery and strain level pathogen characterization using liquid chromatography/mass spectrometry (LC/MS) with electrospray ionization is described. This method was applied to two pathogenic Clostridium species: C. difficile and C. perfringens. Seven marker proteins per species (fourteen total) were successfully implemented to speciate unknowns during a blind study and could enhance serological and genetic approaches by serving as new targets for detection. Two sets of C. perfringens isolates that were 100% similar by pulsed-field gel electrophoresis (PFGE) were distinguished using LC/MS, demonstrating the high specificity of this approach. The use of LC/MS is less labor intensive than PFGE, affords greater specificity than real-time PCR, and requires no primers or antibodies.

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      Affiliations
      [1 ] Commonwealth of Virginia, Division of Consolidated Laboratory Services, 600 N. 5th Street, Richmond, VA 23219, USA.
      Journal
      J. Microbiol. Methods
      Journal of microbiological methods
      Elsevier BV
      1872-8359
      0167-7012
      May 2009
      : 77
      : 2
      19318054 S0167-7012(09)00028-1 10.1016/j.mimet.2009.01.013

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