Development of reagents and assays for the detection of pathogenic Burkholderia species

Omar Qazi; Mridula Rani; Annie J. Gnanam; Thomas W. Cullen; Christopher M. Stead; Haley Kensing; Kate McCaul; Sarah Ngugi; Joann L. Prior; Alexandria Lipka; Judit M. Nagy; Gregory C. Whitlock; Barbara M. Judy; Sarah V. Harding; Richard W. Titball; Sachdev S. Sidhu; M. Stephen Trent; G. Barrie Kitto; Alfredo Torres; D. Mark Estes; Brent Iverson; George Georgiou; Katherine A. Brown

doi:10.1039/C005422B

Development of reagents and assays for the detection of pathogenic Burkholderia species

Omar Qazi,^a Mridula Rani,^a Annie J. Gnanam,^a Thomas W. Cullen,^b Christopher M. Stead,^b Haley Kensing,^a Kate McCaul,^a Sarah Ngugi,^c Joann L. Prior,^c Alexandria Lipka,^d Judit M. Nagy,^e Gregory C. Whitlock,^f Barbara M. Judy,^g Sarah V. Harding,^c Richard W. Titball,^h Sachdev S. Sidhu,ⁱ M. Stephen Trent,^b G. Barrie Kitto,^aj Alfredo Torres,^gk D. Mark Estes,^gk Brent Iverson,^aj George Georgiou^al and Katherine A. Brown*^adj

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* Corresponding authors

^a Institute for Cellular and Molecular Biology, The University of Texas at Austin, 1 University Station A4800, Austin, Texas, USA
E-mail: kate01@mail.utexas.edu
Tel: +1 (512) 471 9483

^b Section of Molecular Genetics and Microbiology, The University of Texas at Austin, Austin, Texas, USA

^c Defence Science and Technology Laboratory, Porton Down, Salisbury, Wiltshire, UK

^d Department of Life Sciences, Imperial College London, Exhibition Road, London, UK

^e Institute for Biomedical Engineering and the Department of Chemistry, Imperial College London, Exhibition Road, London, UK; Deceased

^f Department of Clinical Laboratory Sciences, University of Texas Medical Branch, Galveston, Texas, USA

^g Department of Pathology, University of Texas Medical Branch, Galveston, Texas, USA

^h School of Biosciences, University of Exeter, Geoffrey Pope Building, UK

ⁱ Terence Donnelly Center for Cellular and Biomolecular Research, Banting and Best Department of Biomedical Research, University of Toronto, Toronto, Ontario, Canada

^j Department of Chemistry and Biochemistry, University of Texas at Austin, Austin, Texas, USA

^k Department of Microbiology and Immunology and the Sealy Center for Vaccine Development, University of Texas Medical Branch, Galveston, Texas, USA

^l Department of Chemical Engineering and Biomedical Engineering, The University of Texas at Austin, Austin, Texas, USA

Abstract

Rapid detection of the category B biothreat agents Burkholderia pseudomallei and Burkholderia mallei in acute infections is critical to ensure that appropriate treatment is administered quickly to reduce an otherwise high probability of mortality (ca. 40% for B. pseudomallei). We are developing assays that can be used in clinical laboratories or security applications for the direct detection of surface-localized and secreted macromolecules produced by these organisms. We present our current medium-throughout approach for target selection and production of Burkholderiamacromolecules and describe the generation of a Fab molecule targeted to the B. malleiBimA protein. We also present development of prototype assays for detecting Burkholderia species using anti-lipopolysaccharide antibodies.

This article is part of the themed collection: Analysis for Healthcare Diagnostics and Theranostics

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Development of reagents and assays for the detection of pathogenic Burkholderia species

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