Development and validation of an ELISA for a biomarker of thyroid dysfunction, thyroid peroxidase autoantibodies (TPO-Ab), in dried blood spots
| Autor: | J. Josh Snodgrass, Tara J Cepon-Robins, Maureen J. Devlin, Larry S. Sugiyama, Geeta N. Eick, Paul Kowal |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Male
Physiology lcsh:GN49-298 medicine.disease_cause Autoantigens 0302 clinical medicine Dried blood spots Limit of Detection Iron-Binding Proteins Orthopedics and Sports Medicine 030212 general & internal medicine Dried blood biology Spots Methodology Article Middle Aged Old Friends Hypothesis Dried blood spot Biomarker (medicine) Female Adult Adolescent 030209 endocrinology & metabolism Human Factors and Ergonomics Enzyme-Linked Immunosorbent Assay Thyroid peroxidase autoantibodies Iodide Peroxidase 03 medical and health sciences Young Adult Thyroid peroxidase Thyroid dysfunction Physiology (medical) medicine Humans Autoantibodies lcsh:Physical anthropology. Somatology business.industry Public Health Environmental and Occupational Health Autoantibody Reproducibility of Results Immune dysregulation Thyroid Diseases Anthropology Immunology biology.protein Linear Models Dried Blood Spot Testing business In-house immunoassay Biomarkers |
| Zdroj: | Journal of Physiological Anthropology Journal of Physiological Anthropology, Vol 39, Iss 1, Pp 1-8 (2020) |
| ISSN: | 1880-6805 1880-6791 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full text from SpringerLink