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Clinical and Vaccine Immunology, May 2009, p. 765-771, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00006-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Competitive Electrochemiluminescence Wash and No-Wash Immunoassays for Detection of Serum Antibodies to Smooth Brucella Strains{triangledown}

Iain Thompson,1 John McGiven,1* Jason Sawyer,2 Rachel Thirlwall,1 Nicola Commander,1 and Judy Stack1

Brucella Research Group, Statutory and Exotic Bacteria, Veterinary Laboratories Agency (VLA), Woodham Lane, New Haw, Surrey KT15 3NB, United Kingdom,1 Technology Transfer Unit, Biotechnology Department, Veterinary Laboratories Agency (VLA), Woodham Lane, New Haw, Surrey KT15 3NB, United Kingdom2

Received 7 January 2009/ Returned for modification 28 January 2009/ Accepted 25 February 2009

Brucellosis is a bacterial zoonotic disease of major global importance. Natural hosts for Brucella species include animals of economic significance, such as cattle and small ruminants. Controlling brucellosis in natural hosts by high-throughput serological testing followed by the slaughter of seropositive animals helps to prevent disease transmission. This study aimed to convert an existing competitive enzyme-linked immunosorbent assay (cELISA), used for the serodiagnosis of brucellosis in ruminants, to two electrochemiluminescence (ECL) immunoassays on the Meso Scale Discovery (MSD) platform. The first assay employed a conventional plate washing step as part of the protocol. The second was a no-wash assay, made possible by the proximity-based nature of ECL signal generation by the MSD platform. Both ECL wash and no-wash assays closely matched the parent cELISA for diagnostic sensitivity and specificity. The results also demonstrated that both ECL assays met World Organization for Animal Health (OIE) standards, as defined by results for the OIE standard serum (OIEELISASPSS). This report is the first to describe an ECL assay incorporating lipopolysaccharide, an ECL assay for serodiagnosis of a bacterial infectious disease, a separation-free (no-wash) ECL assay for the detection of serum antibodies, and the use of the MSD platform for serodiagnosis. The simple conversion of the cELISA to the MSD platform suggests that many other serodiagnostic tests could readily be converted. Furthermore, the alignment of these results with the multiplex capability of the MSD platform offers the potential of no-wash multiplex assays to screen for several diseases.

* Corresponding author. Mailing address: Brucella Research Group, Veterinary Laboratories Agency, New Haw, Surrey KT15 3NB, United Kingdom. Phone: 44 1932 357227. Fax: 44 1932 357875. E-mail: j.mcgiven{at}vla.defra.gsi.gov.uk

{triangledown} Published ahead of print on 4 March 2009.

Clinical and Vaccine Immunology, May 2009, p. 765-771, Vol. 16, No. 5
1071-412X/09/$08.00+0     doi:10.1128/CVI.00006-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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