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Journal of Clinical Microbiology, May 2009, p. 1436-1442, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.02380-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Rapid Determination of Quinolone Resistance in Acinetobacter spp.{triangledown}

Kristine M. Hujer,1,2 Andrea M. Hujer,2 Andrea Endimiani,1,2 Jodi M. Thomson,3 Mark D. Adams,4 Karrie Goglin,4 Philip N. Rather,5 Thuy-Trang D. Pennella,6 Christian Massire,6 Mark W. Eshoo,6 Rangarajan Sampath,6 Lawrence B. Blyn,6 David J. Ecker,6* and Robert A. Bonomo1,2,3,7*

Department of Medicine, Case Western Reserve University School of Medicine, Cleveland, Ohio,1 Research Service, Louis Stokes Cleveland Department of Veterans Affairs Medical Center, Cleveland, Ohio,2 Departments of Pharmacology,3 Genetics, Case Western Reserve University School of Medicine, Cleveland, Ohio,4 Departments of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia,5 Ibis Biosciences, Carlsbad, California,6 Molecular Biology and Microbiology, Case Western Reserve University School of Medicine, Cleveland, Ohio7

Received 11 December 2008/ Returned for modification 3 February 2009/ Accepted 11 March 2009

In the treatment of serious bacterial infections, the rapid institution of appropriate antimicrobial chemotherapy may be lifesaving. Choosing the correct antibiotic or combination of antibiotics is becoming very important, as multidrug resistance is found in many pathogens. Using a collection of 75 well-characterized multidrug-resistant (MDR) Acinetobacter sp. isolates, we show that PCR followed by electrospray ionization mass spectrometry (PCR/ESI-MS) and base composition analysis of PCR amplification products can quickly and accurately identify quinolone resistance mediated by mutations in the quinolone resistance-determining regions of gyrA and parC, two essential housekeeping genes. Single point mutations detected by PCR/ESI-MS in parC (found in 55/75 of the isolates) and in gyrA (found in 66/75 of the isolates) correlated with susceptibility testing and sequencing. By targeting resistance determinants that are encoded by genes with highly conserved DNA sequences (e.g., gyrA and parC), we demonstrate that PCR/ESI-MS can provide critical information for resistance determinant identification and can inform therapeutic decision making in the treatment of Acinetobacter sp. infections.

* Corresponding author. Mailing address for David J. Ecker: Ibis Biosciences, 1891 Rutherford Road, Carlsbad, CA 92008. Phone: (760) 603-2347. Fax: (760) 603-4653. E-mail: DEcker{at}ibisbio.com. Mailing address for Robert A. Bonomo: Louis Stokes Cleveland Department of Veterans Affairs Medical Center, 10701 East Blvd., Cleveland, OH 44106. Phone: (216) 791-3800, ext. 4399. Fax: (216) 229-8509. E-mail: robert.bonomo{at}med.va.gov

{triangledown} Published ahead of print on 18 March 2009.

Journal of Clinical Microbiology, May 2009, p. 1436-1442, Vol. 47, No. 5
0095-1137/09/$08.00+0     doi:10.1128/JCM.02380-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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