- Title
- Development and validation of an ELISA for a biomarker of thyroid dysfunction, thyroid peroxidase autoantibodies (TPO-Ab), in dried blood spots
- Creator
- Eick, Geeta N.; Cepon-Robins, Tara J.; Devlin, Maureen J.; Kowal, Paul; Sugiyama, Larry S.; Snodgrass, J. Josh
- Relation
- Journal of Physiological Anthropology Vol. 39, Issue 1, no. 16
- Publisher Link
- http://dx.doi.org/10.1186/s40101-020-00228-8
- Publisher
- BioMed Central (BMC)
- Resource Type
- journal article
- Date
- 2020
- Description
- Background: The prevalence of allergic and autoimmune conditions has been steadily increasing in wealthy nations over the past century. One hypothesis put forward to explain this is the Old Friends Hypothesis, which posits that increased hygiene, urbanization, and lifestyle changes have reduced our exposure to parasites and microbes that we co-evolved with, resulting in immune dysregulation. However, research in traditionally living populations, who are exposed to greater parasite and pathogen loads such as those encountered during our evolution, is limited, in part due to a lack of minimally invasive, field-friendly biomarkers of autoimmune disorders. We therefore developed an ELISA to assess positivity for thyroid peroxidase autoantibody (TPO-Ab), an indicator of autoimmune thyroid disease, based on dried blood spot (DBS) samples. Results: We used the Accubind anti-thyroid peroxidase test system to screen our validation samples comprising matched fingerprick DBS, venous DBS, and plasma samples from 182 adults. After confirming that we had TPO-Ab-positive individuals in our validation sample (n = 12), we developed an indirect ELISA to measure TPO-Ab levels from one 3-mm DBS punch. The sensitivity and specificity of our assay for DBS samples ranged from 91.7–100% and 98.2–98.8%, respectively, using a cut-off value of ≥ 26 IU/mL. Intra-assay reliability for duplicate quality control DBS punches was 5.2%, while inter-assay reliability ranged from 11.5–24.4% for high, medium, and low DBS controls. Dilutional linearity ranged from 80 to 120%, and spike and recovery experiments indicated that the DBS matrix does not interfere with the detection of TPO-Ab. TPO-Ab levels remained stable in DBS samples stored at − 28 °C or − 80 °C, but decreased over time in DBS samples kept at 22 °C or at 37 °C. Conclusions: We developed an in-house, kit-independent indirect ELISA assay to determine individuals’ TPO-Ab positivity based on dried blood spots, representing a cost-effective method with potential applications in a range of research settings.
- Subject
- in-house immunoassay; thyroid peroxidase autoantibodies; dried blood spots; old friends hypothesis
- Identifier
- http://hdl.handle.net/1959.13/1433015
- Identifier
- uon:39149
- Identifier
- ISSN:1880-6805
- Rights
- © The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
- Language
- eng
- Full Text
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