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Clinical and Vaccine Immunology, June 2007, p. 665-668, Vol. 14, No. 6
1071-412X/07/$08.00+0     doi:10.1128/CVI.00480-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evaluation of an Enzyme Immunoassay for Detection of Immunoglobulin M Antibodies to West Nile Virus and the Importance of Background Subtraction in Detecting Nonspecific Reactivity{triangledown}

Mindy L. Rawlins,1* Erica M. Swenson,1 Harry R. Hill,1,2 and Christine M. Litwin1,2

Associated Regional and University Pathologists Institute for Clinical and Experimental Pathology,1 Department of Pathology, Pediatrics and Medicine, University of Utah School of Medicine, Salt Lake City, Utah2

Received 19 December 2006/ Returned for modification 31 January 2007/ Accepted 27 March 2007

Since the introduction of West Nile virus (WNV) in the United States in 1999, several assays have become commercially available to detect antibodies against WNV. Capture enzyme-linked immunosorbent assays (ELISAs) for the detection of WNV-specific immunoglobulin M (IgM) have been approved by the Food and Drug Administration for clinical testing and are available from Focus Diagnostics and PanBio, Inc. The Focus Diagnostics IgM capture ELISA utilizes a background subtraction protocol in order to detect nonspecific reactivity due to rheumatoid factor, heterophile antibodies, or other interfering substances. A background subtraction procedure is not currently recommended for the PanBio IgM capture ELISA. In previous experiments, we determined the agreement, sensitivity, and specificity of the PanBio first-generation IgM capture ELISA compared to an immunofluorescence assay and the Centers for Disease Control and Prevention's IgM capture ELISA. The PanBio assay has since been reformulated to improve the specificity of the assay. We evaluated the reformulated PanBio assay with and without an antigen subtraction procedure and compared the results to the Focus IgM capture ELISA. Agreement, sensitivity, and specificity of the PanBio assay were, respectively, 85%, 95%, and 76% without the subtraction protocol and 94%, 95%, and 93% with the subtraction protocol. In general, when the subtraction protocol was applied to the PanBio IgM capture ELISA, there was a reduction in some, but not all, false-positive results. We suggest that all WNV IgM assays be standardized with a procedure such as background subtraction to eliminate nonspecific reactivity that may cause false-positive results.

* Corresponding author. Mailing address: ARUP Institute for Clinical and Experimental Pathology, 500 Chipeta Way, Salt Lake City, UT 84108. Phone: (801) 583-2787, ext. 2840. Fax: (801) 584-5109. E-mail: mindy.rawlins{at}aruplab.com

{triangledown} Published ahead of print on 11 April 2007.

Clinical and Vaccine Immunology, June 2007, p. 665-668, Vol. 14, No. 6
1071-412X/07/$08.00+0     doi:10.1128/CVI.00480-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Prince, H. E., Yeh, C., Lape-Nixon, M. (2008). Development of a More Efficient Algorithm for Identifying False-Positive Reactivity Results in a Dengue Virus Immunoglobulin M Screening Assay. CVI 15: 1304-1306 [Abstract] [Full Text]  


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