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Clinical and Diagnostic Laboratory Immunology, March 2004, p. 313-319, Vol. 11, No. 2
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.2.313-319.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Combined Use of Enzyme-Linked Immunosorbent Assay and Flow Cytometry To Detect Antibodies to Trypanosoma cruzi in Domestic Canines in Texas

Sean V. Shadomy, Stephen C. Waring, and Cynthia L. Chappell*

Center for Infectious Diseases, The University of Texas Health Science Center at Houston School of Public Health, Houston, Texas 77030

Received 10 September 2003/ Returned for modification 21 October 2003/ Accepted 25 November 2003

Canines may be sentinels and/or reservoirs for human Trypanosoma cruzi exposures. This study adapted a method originally designed for human diagnostics to detect serum immunoglobulin G to T. cruzi in canines. The method combined an enzyme-linked immunosorbent assay (ELISA) for screening and flow cytometry detection of anti-live trypomastigote antibodies (ALTA) for confirmation. The assays were optimized by using known positive and negative control canine sera, and cutoff values were established. The ELISA and ALTA assay easily distinguished between reactive (positive controls) and nonreactive (negative controls) sera and were used to test sera collected in a cross-sectional seroprevalence survey of 356 domestic canines from Harris County, Tex., and the surrounding area. Fifty-three (14.9%) of 356 asymptomatic canines in the survey were positive by ELISA, and 5 (1.4%) were confirmed positive with the ALTA assay, with an additional 4 (1.1%) canines classified as "suspect positive." Thus, the overall prevalence of T. cruzi antibodies in this population was 2.6%. This is the first U.S. study to use the combination of ELISA and ALTA to detect serum antibodies to T. cruzi and the first report of the prevalence of T. cruzi infection in domestic canines in the Houston, Tex. (Harris County), region. Our results demonstrate that the combination of ELISA and ALTA has been successfully adapted for use in testing canines for serological evidence of T. cruzi infection. Seroprevalence survey results suggest that T. cruzi antibody-positive domestic canines in the peridomestic setting are present in the Houston, Tex., region and further suggest that T. cruzi is enzootic in the region.

* Corresponding author. Present address: University of Texas School of Public Health, 1200 Herman Pressler Dr., Ste. 118A, Houston, TX 77030. Phone: (713) 500-9026. Fax: (713) 500-9020. E-mail: Cynthia.L.Chappell{at}uth.tmc.edu.

Clinical and Diagnostic Laboratory Immunology, March 2004, p. 313-319, Vol. 11, No. 2
1071-412X/04/$08.00+0     DOI: 10.1128/CDLI.11.2.313-319.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.





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