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Clinical and Vaccine Immunology, December 2008, p. 1834-1838, Vol. 15, No. 12
1071-412X/08/$08.00+0     doi:10.1128/CVI.00238-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Multiplex Immunoassay for Serological Diagnosis of Mycobacterium bovis Infection in Cattle{triangledown}

Clare Whelan,1 Eduard Shuralev,1 Grainne O'Keeffe,1 Paula Hyland,1 Hang Fai Kwok,2 Philip Snoddy,2 Amanda O'Brien,1 Marie Connolly,1 Padraig Quinn,1 Matt Groll,3 Todd Watterson,3 Sara Call,3 Kevin Kenny,4 Anthony Duignan,5 Mary Jo Hamilton,6 Bryce M. Buddle,7 James A. Johnston,2 William C. Davis,6 Shane A. Olwill,2 and John Clarke1*

Enfer Scientific, Unit T, M7 Business Park, Newhall, Naas, County Kildare, Ireland,1 Fusion Antibodies Ltd., Unit 4, Springbank Industrial Estate, Pembroke Loop Rd., BT17 0QL, Northern Ireland,2 Quansys Biosciences, 365 North 600 West, Logan, Utah 84321,3 Central Veterinary Research Laboratory, Backweston, County Dublin, Ireland,4 Department of Agriculture and Food, Kildare St., Dublin 2, Ireland,5 Department of Veterinary Microbiology and Pathology, College of Veterinary Medicine, Washington State University, Pullman, Washington 99164-7040,6 AgResearch, Hopkirk Research Institute, Palmerston North, New Zealand7

Received 27 June 2008/ Returned for modification 18 July 2008/ Accepted 4 October 2008

Efforts to develop a better diagnostic assay for bovine tuberculosis have shown that the sensitivity and specificity of an assay can be improved by the use of two or more antigens. As reported here, we developed a multiplex chemiluminescent immunoassay that can simultaneously detect antibody activity to 25 antigens in a single well in a 96-well plate array format. The chemiluminescent signal is captured with a digital imaging system and analyzed with a macro program that tracks each serum for its pattern of antibody activity for Mycobacterium bovis antigens. The comparison of sera from 522 infected and 1,489 uninfected animals showed that a sensitivity of 93.1% and a specificity of 98.4% can be achieved with a combination of antigens. The assay system is rapid and can be automated for use in a centralized laboratory.

* Corresponding author. Mailing address: Enfer Scientific, Unit T, M7 Business Park, Newhall, Naas, County Kildare, Ireland. Phone: (353) 45 983800. Fax: (353) 45 983801. E-mail: johnclarke{at}enfergroup.com

{triangledown} Published ahead of print on 15 October 2008.

Clinical and Vaccine Immunology, December 2008, p. 1834-1838, Vol. 15, No. 12
1071-412X/08/$08.00+0     doi:10.1128/CVI.00238-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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