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LcrV Capture Enzyme-Linked Immunosorbent Assay for Detection of Yersinia pestis from Human Samples

Maria J. C. Gomes-Solecki,^1, Anne G. Savitt,² Rebecca Rowehl,² John D. Glass,³ James B. Bliska,^1,2 and Raymond J. Dattwyler^1*

Center for Infectious Diseases,¹ Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook,² Brookhaven National Laboratory, Upton, New York³

Received 20 August 2004/ Returned for modification 24 September 2004/ Accepted 1 November 2004

In the United States, there is currently a major gap in the diagnostic capabilities with regard to plague. To address this, we developed an antigen capture assay using an essential virulence factor secreted by Yersinia spp., LcrV, as the target antigen. We generated anti-LcrV monoclonal antibodies (MAbs) and screened them for the ability to bind bacterially secreted native Yersinia pestis LcrV. Anti-LcrV MAb 19.31 was used as a capture antibody, and biotinylated MAb 40.1 was used for detection. The detection limit of this highly sensitive Yersinia LcrV capture enzyme-linked immunosorbent assay is 0.1 ng/ml. The assay detected LcrV from human sputum and blood samples treated with concentrations as low as 0.5 ng/ml of bacterially secreted native Y. pestis LcrV. This assay could be used as a tool to help confirm the diagnosis of plague in patients presenting with pneumonia.

Present address: Department of Microbiology and Immunology, New York Medical College, Valhalla, NY 10595.