Tmrna â a novel high-copy-number rna diagnostic target â its application forstaphylococcus aureusdetection using real-time nasba
O'Grady, Justin Lacey, Katrina Glynn, Barry Smith, Terry J. Barry, Thomas Maher, Majella
O'Grady, Justin
Lacey, Katrina
Glynn, Barry
Smith, Terry J.
Barry, Thomas
Maher, Majella
Publication Date
2009-12-01
Type
Article
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Citation
O'Grady, Justin; Lacey, Katrina; Glynn, Barry; Smith, Terry J. Barry, Thomas; Maher, Majella (2009). Tmrna â a novel high-copy-number rna diagnostic target â its application forstaphylococcus aureusdetection using real-time nasba. FEMS Microbiology Letters 301 (2), 218-223
Abstract
A real-time nucleic acid sequence-based amplification assay, targeting tmRNA, was designed for the rapid identification of Staphylococcus aureus. The selectivity of the assay was confirmed against a panel of 76 Staphylococcus strains and species and 22 other bacterial species. A detection limit of 1 cell equivalent was determined for the assay. A chimeric in vitro transcribed internal amplification control was developed and included in the assay. Application of the assay in natural and artificially contaminated unpasteurized (raw) milk enabled detection of 1-10 CFUS. aureus mL-1 in 3-4 h, without the need for culture enrichment. Staphylococcus aureus was detected in all artificially contaminated milk samples (n=20) and none of the natural milk samples (n=20). Microbiological analysis of the natural milk samples was performed in parallel according to ISO 6888-3 and confirmed the absence of S. aureus. The method developed in this study has the potential to enable the specific detection of S. aureus in raw milk in a significantly shorter time frame than current standard methods. The assay further demonstrates the usefulness of tmRNA/ssrA as a nucleic acid diagnostic target.
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Publisher
Oxford University Press (OUP)
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Attribution-NonCommercial-NoDerivs 3.0 Ireland