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Dot enzyme-linked immunosorbent assay (ELISA) for the detection of Toxocara infection using a rat model

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Abstract

Toxocariasis is a zoonotic disease usually caused by dog and cat roundworms, Toxocara canis and T. cati. Detection and diagnosis is difficult in paratenic and accidental hosts, including humans, as they cannot be detected through conventional methods such as fecal examination. Diagnosis therefore relies on immunological methods and molecular methods such as enzyme-linked immunosorbent assay (ELISA) and Western Blot, which are both time-consuming and requires sophisticated equipment. In the Philippines, only a few studies are available on Toxocara seroprevalence. Therefore, there is a need to adapt methods for serodiagnosis of Toxocara infection in humans for the Philippine setting. A dot enzyme linked immunosorbent assay (dot-ELISA) was standardized using T. canis excretory-secretory antigens. Test sera were collected from laboratory rats (Sprague–Dawley strain) experimentally infected with embryonated eggs of T. canis and Ascaris suum as well as rice field rats naturally infected with Taenia taeniaeformis and Nippostrongylus sp. Optimum conditions used were 20 µg/ml antigen concentration and 1:10 serum dilution. The sensitivity, specificity, positive, and negative predictive values were 90% (95% CI 55.5–99.7%), 100% (95% CI 69.2–100.0%), 100% (95% CI 66.4–100%), and 90.9% (95% CI 58.7–99.8%), respectively. Dot-ELISA has the potential to be developed as a cheaper, simpler, and more practical method for detection of anti-Toxocara antibodies on accidental hosts. This is a preliminary study conducted on experimental animals before optimization and standardization for human serum samples.

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Acknowledgements

The researchers would like to thank the National Immunological Testing Laboratory (NITL), the National Institute of Molecular Biology and Biotechnology (BIOTECH) for the assistance and facilities provided. This research was supported by the University of the Philippines Enhanced Work and Research Grant (UP ECWRG) and the Department of Science and Technology – Accelerated Science and Technology Human Resource and Development Program (DOST-ASTHRDP).

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Authors and Affiliations

  1. Institute of Biological Sciences, College of Arts and Sciences, University of the Philippines Los Baños, Los Baños, Laguna, Philippines

    Vachel Gay V. Paller, Cyrelle M. Besana & Isabel Kristine M. Valdez

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  1. Vachel Gay V. Paller
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  2. Cyrelle M. Besana
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  3. Isabel Kristine M. Valdez
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Contributions

Conceived and designed the experiments: VGVP, IKMV; Performed the experiments: IKMV, CMB; Data analysis: IKMV; Contributed reagents/materials/analysis tools: VGVP, IKMV, CMB; All authors participated in writing the final paper.

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Correspondence to Vachel Gay V. Paller.

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Paller, V.G.V., Besana, C.M. & Valdez, I.K.M. Dot enzyme-linked immunosorbent assay (ELISA) for the detection of Toxocara infection using a rat model. J Parasit Dis 41, 933–939 (2017). https://doi.org/10.1007/s12639-017-0914-6

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  • DOI: https://doi.org/10.1007/s12639-017-0914-6

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