CVI Accepts, published online ahead of print on 4 March 2009

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Clin. Vaccine Immunol. doi:10.1128/CVI.00006-09
Copyright (c) 2009, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Competitive electrochemiluminescence (ECL) wash and no-wash immunoassays for the detection of serum antibodies to smooth Brucella strains

Iain Thompson, John McGiven^*, Jason Sawyer, Rachel Thirlwall, Nicola Commander, and Judy Stack

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

^* To whom correspondence should be addressed. Email: j.mcgiven{at}vla.defra.gsi.gov.uk.

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 sero-positive animals helps 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 Organisation for Animal Health (OIE) standards as defined by results of the OIE Standard Sera (OIEELISA_SPSS).

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 be readily 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.