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Use of Resequencing Oligonucleotide Microarrays for Identification of Streptococcus pyogenes and Associated Antibiotic Resistance Determinants

Louis Davignon,¹ Elizabeth A. Walter,^2,3 Kate M. Mueller,⁴ Christopher P. Barrozo,⁵ David A. Stenger,⁶ Baochuan Lin,^6* on behalf of the Epidemic Outbreak Surveillance Consortium,

Malcolm Grow Medical Center, Andrews Air Force Base, Maryland 20762,¹ Epidemic Outbreak Surveillance Advanced Diagnostics Laboratory, Lackland Air Force Base, San Antonio, Texas 78236,² Texas A&M University Systems, San Antonio, Texas 78223,³ Nova Research Inc., Alexandria, VA 22308,⁴ DoD Center for Deployment Health Research, Naval Health Research Center, San Diego, California 92152,⁵ Center for Bio/Molecular Science & Engineering, Code 6900, Naval Research Laboratory, Washington, DC 20375⁶

Received 25 February 2005/ Returned for modification 19 May 2005/ Accepted 15 August 2005

Group A streptococci (GAS) are responsible for a wide variety of human infections associated with considerable morbidity and mortality. Ever since the first systematic effort by Lancefield to group Streptococcus species by M protein variants, the detection and characterization of Streptococcus by different methods have been an evolving process. The ideal assay for GAS identification not only would provide quick and accurate diagnostic results but also would reveal antibiotic resistance patterns and genotype information, aiding not only in treatment but in epidemiologic assessment as well. The oligonucleotide microarray is a promising new technology which could potentially address this need. In this study, we evaluated the usefulness of oligonucleotide resequencing microarrays for identifying GAS and its associated antibiotic resistance markers. We demonstrated an assay platform that combines the use of resequencing DNA microarrays with either random nucleic acid amplification or multiplex PCR for GAS detection. When detecting Streptococcus pyogenes from coded clinical samples, this approach demonstrated an excellent concordance with a more established culture method. To this end, we showed the potential of resequencing microarrays for efficient and accurate detection of GAS and its associated antibiotic resistance markers with the benefit of sequencing information from microarray analysis.

Supplemental material for this article may be found at http://jcm.asm.org/.

The Epidemic Outbreak Surveillance Consortium is an Air Force Medical Service initiative. The following are participating members of the EOS Consortium: (i) sponsorship, Peter F. Demitry (USAF/SGR) and Theresa Lynn Difato (USAF/SGR); (ii) executive board and principal investigators, Eric H. Hanson (The George Washington University [IPA]), Rosana R. Holliday (USAF/SGR [Ctr]), Robb K. Rowley (The George Washington University [IPA]), and Clark Tibbetts (The George Washington University [IPA]); (iii) operational board and senior scientists, Brian K. Agan (Wilford Hall Medical Center), Jerry Diao (USAF/SGR [Ctr]), Russell P. Kruzelock (Virginia Tech [IPA]), David A. Stenger (Naval Research Laboratory), and Elizabeth A. Walter (Texas A&M University Systems San Antonio [IPA]); (iv) technical advisors and collaborating investigators, Luke Daum (Air Force Institute for Operational Health), David Metzgar (Navy Health Research Center), Debra Niemeyer (USAF/SGR), and Kevin Russell (Navy Health Research Center); (v) research and clinical staff, Marie J. Archer (Naval Research Laboratory), Roger Bravo (Lackland AFB, Tex.), Nikki Freed (Naval Health Research Center), Julie Fuller (Naval Health Research Center), John Gomez (Lackland AFB, Tex.), Kevin Gratwick (Naval Health Research Center), Michael Jenkins (Wilford Hall Medical Center), Margaret Jesse (Lackland AFB, Tex.), Barry Johnson (Lackland AFB, Tex.), Erin Lawrence (Naval Health Research Center), Baochuan Lin (Naval Research Laboratory), Carolyn E. Meador (NOVA Research Incorporated), Hernan Melgarejo (Lackland AFB, Tex.), Kate M. Mueller (NOVA Research Incorporated), Chris Olsen (USAF/SGR [Ctr]), David Pearson (Lackland AFB, Tex.), Anjan Purkayastha (USAF/SGR [Ctr]), Jose J. Santiago (Lackland AFB, Tex.), Donald Seto (George Mason University [IPA]), Francine Fuentes Stotler (Lackland AFB, Tex.), Dzung Thach (Naval Research Laboratory), Jennifer A. Thornton (NOVA Research Incorporated), Zheng Wang (Naval Research Laboratory), Daisy Watson (Lackland AFB, Tex.), Sue A. Worthy (Lackland AFB, Tex.), and Gary J. Vora (Naval Research Laboratory); and (vi) operations support staff, Kenya Grant (USAF/SGR [Ctr]), Cheryl J. James (USAF/SGR [Ctr]), and Kathy Word (USAF/SGR [Ctr]).

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