Rachel Busselman


Curriculum vitae



Ecology and Evolutionary Biology

Texas A&M University

Ecology and Evolutionary Biology
School of Veterinary Medicine
Department of Veterinary Integrative Biosciences



Validation of a multiplex microsphere immunoassay for detection of antibodies to Trypanosoma cruzi in dogs


Journal article


Carlos A. Rodriguez, Rachel E. Busselman, Huifeng Shen, Ashley B. Saunders, Rick Tarleton, S. Hamer
Journal of Veterinary Diagnostic Investigation, 2023

Semantic Scholar DOI PubMed
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APA   Click to copy
Rodriguez, C. A., Busselman, R. E., Shen, H., Saunders, A. B., Tarleton, R., & Hamer, S. (2023). Validation of a multiplex microsphere immunoassay for detection of antibodies to Trypanosoma cruzi in dogs. Journal of Veterinary Diagnostic Investigation.


Chicago/Turabian   Click to copy
Rodriguez, Carlos A., Rachel E. Busselman, Huifeng Shen, Ashley B. Saunders, Rick Tarleton, and S. Hamer. “Validation of a Multiplex Microsphere Immunoassay for Detection of Antibodies to Trypanosoma Cruzi in Dogs.” Journal of Veterinary Diagnostic Investigation (2023).


MLA   Click to copy
Rodriguez, Carlos A., et al. “Validation of a Multiplex Microsphere Immunoassay for Detection of Antibodies to Trypanosoma Cruzi in Dogs.” Journal of Veterinary Diagnostic Investigation, 2023.


BibTeX   Click to copy

@article{carlos2023a,
  title = {Validation of a multiplex microsphere immunoassay for detection of antibodies to Trypanosoma cruzi in dogs},
  year = {2023},
  journal = {Journal of Veterinary Diagnostic Investigation},
  author = {Rodriguez, Carlos A. and Busselman, Rachel E. and Shen, Huifeng and Saunders, Ashley B. and Tarleton, Rick and Hamer, S.}
}

Abstract

The vector-borne protozoan parasite Trypanosoma cruzi causes Chagas disease in humans, dogs, and many other mammalian hosts. Canine Chagas disease is increasingly diagnosed in dogs of the southern United States where triatomine insect vectors occur, and there are limited veterinary testing options; only the indirect fluorescent antibody (IFA) test is offered at a single accredited diagnostic laboratory. We evaluated a multiplex microsphere immunoassay (MIA) for the detection of antibodies against T. cruzi in dogs and compared it with existing serologic methods to establish cutoff values and relative sensitivity and specificity. We tested 135 canine sera that had been characterized using the IFA and off-label use of 2 commercial rapid assays with our multiplex MIA against 12 antigens: 9 T. cruzi antigens, a negative control recombinant protein (green fluorescent protein, GFP), a Leishmania antigen, and a canine parvovirus antigen (used as an antibody control given near-ubiquitous parvoviral vaccination). The median fluorescence intensity (MFI) ratio between each T. cruzi antigen and GFP was calculated for every sample. Samples with an antigen:GFP MFI ratio > 4 SDs above the mean of 25 known-negative sera were considered positive to that antigen. Samples testing positive to ≥ 2 antigens were considered positive for T. cruzi antibodies. Compared to the IFA, our multiplex MIA had a relative sensitivity of 100% and specificity of 97.0%. Given its precision, high-throughput format, potential for automation, and lack of subjective interpretation, our multiplex MIA should be considered a valid and improved assay for T. cruzi antibodies in dogs.


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