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Identification of Staphylococcus aureus and Determination of Methicillin Resistance Directly from Positive Blood Cultures by Isothermal Amplification and a Disposable Detection Device

BioHelix Corporation, Beverly, Massachusetts 01915,¹ Departments of Medicine,² Pathology, Vanderbilt University Medical Center, Nashville, Tennessee 37232,³ Lahey Clinic Medical Center, Burlington, Massachusetts 01805⁴

Received 19 November 2007/ Returned for modification 4 January 2008/ Accepted 16 January 2008

A simple, rapid, and user-friendly procedure has been developed to identify Staphylococcus aureus and determine its methicillin resistance directly from gram-positive cocci in cluster-containing blood culture medium. The specimens were diluted and heated prior to amplification of the nuc and mecA genes with isothermal helicase-dependent amplification. Amplicons were detected using a disposable detection device. The analytical sensitivity of the assays was 50 CFU per reaction, and the clinical sensitivity and specificity were both 100% for S. aureus detection and 100% and 98% for methicillin resistance determination, respectively.

Published ahead of print on 30 January 2008.