1887

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Volume 65, Issue 1

Single fluorophore melting curve analysis for detection of hypervirulent Free

Abstract

This study demonstrates a novel detection assay able to identify and subtype strains of . Primers carefully designed for melting curve analysis amplify DNA from three genes, , and , during quantitative (q)PCR. The gene allows for confirmation of organism presence, whilst the and genes allow for differentiation of virulence status, as deletions in the gene and the concurrent presence of the gene, which produces binary toxin, are associated with hypervirulence. Following qPCR, subtyping is then achieved by automated, inline melting curve analysis using only a single intercalating dye and verified by microchip electrophoresis. This assay represents a novel means of distinguishing between toxigenic and hypervirulent strains NAP1/027/BI and 078 ribotype, which are highly prevalent hypervirulent strains in humans. This methodology can help rapidly detect and identify strains that impose a significant health and economic burden in hospitals and other healthcare settings.

  • Received:
  • Accepted:
  • Published Online:
© 2016 The Authors
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2016-01-01
2024-04-19
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Single fluorophore melting curve analysis for detection of hypervirulent Clostridium difficile
J. Med. Microbiol. 65, 62 (2016); https://doi.org/10.1099/jmm.0.000199
/content/journal/jmm/10.1099/jmm.0.000199
/content/journal/jmm/10.1099/jmm.0.000199
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