{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,18]],"date-time":"2026-07-18T01:35:39Z","timestamp":1784338539538,"version":"3.55.0"},"reference-count":29,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,7,16]],"date-time":"2020-07-16T00:00:00Z","timestamp":1594857600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,7,16]],"date-time":"2020-07-16T00:00:00Z","timestamp":1594857600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000001","name":"National Science Foundation of the USA","doi-asserted-by":"crossref","award":["BCS-1638786"],"award-info":[{"award-number":["BCS-1638786"]}],"id":[{"id":"10.13039\/100000001","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100011348","name":"University of Oregon","doi-asserted-by":"crossref","award":["Richard A Bray Fellowship"],"award-info":[{"award-number":["Richard A Bray Fellowship"]}],"id":[{"id":"10.13039\/100011348","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["J Physiol Anthropol"],"published-print":{"date-parts":[[2020,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec>\n<jats:title>Background<\/jats:title>\n<jats:p>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.<\/jats:p>\n<\/jats:sec><jats:sec>\n<jats:title>Results<\/jats:title>\n<jats:p>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 (<jats:italic>n<\/jats:italic> = 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\u2013100% and 98.2\u201398.8%, respectively, using a cut-off value of \u2265 26\u2009IU\/mL. Intra-assay reliability for duplicate quality control DBS punches was 5.2%, while inter-assay reliability ranged from 11.5\u201324.4% 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 \u2212 28\u2009\u00b0C or \u2212 80\u2009\u00b0C, but decreased over time in DBS samples kept at 22\u2009\u00b0C or at 37\u2009\u00b0C.<\/jats:p>\n<\/jats:sec><jats:sec>\n<jats:title>Conclusions<\/jats:title>\n<jats:p>We developed an in-house, kit-independent indirect ELISA assay to determine individuals\u2019 TPO-Ab positivity based on dried blood spots, representing a cost-effective method with potential applications in a range of research settings.<\/jats:p>\n<\/jats:sec>","DOI":"10.1186\/s40101-020-00228-8","type":"journal-article","created":{"date-parts":[[2020,7,16]],"date-time":"2020-07-16T10:03:28Z","timestamp":1594893808000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Development and validation of an ELISA for a biomarker of thyroid dysfunction, thyroid peroxidase autoantibodies (TPO-Ab), in dried blood spots"],"prefix":"10.1186","volume":"39","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7512-3265","authenticated-orcid":false,"given":"Geeta N.","family":"Eick","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tara J.","family":"Cepon-Robins","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maureen J.","family":"Devlin","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul","family":"Kowal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Larry S.","family":"Sugiyama","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. Josh","family":"Snodgrass","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,7,16]]},"reference":[{"key":"228_CR1","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1177\/1757913916650225","volume":"136","author":"SF Bloomfield","year":"2016","unstructured":"Bloomfield SF, Rook GAW, Scott EA, Shanahan F, Stanwell-Smith R, Turner P. Time to abandon the hygiene hypothesis: new perspectives on allergic disease, the human microbiome, infectious disease prevention and the role of targeted hygiene. Perspect Public Health. 2016;136:213\u201324.","journal-title":"Perspect Public Health"},{"key":"228_CR2","first-page":"173","volume":"7","author":"GAW Rook","year":"2012","unstructured":"Rook GAW. A Darwinian view of the hygiene or \u201cold friends\u201d hypothesis. Microbe. 2012;7:173\u201380.","journal-title":"Microbe."},{"key":"228_CR3","volume-title":"Social change, parasite exposure, and immune dysregulation among Shuar forager-horticulturalists of Amazonia: a biocultural case-study in evolutionary medicine","author":"T Robins","year":"2015","unstructured":"Robins T. Social change, parasite exposure, and immune dysregulation among Shuar forager-horticulturalists of Amazonia: a biocultural case-study in evolutionary medicine; 2015."},{"key":"228_CR4","doi-asserted-by":"publisher","first-page":"729","DOI":"10.1210\/jc.2007-1644","volume":"93","author":"A Kondrashova","year":"2008","unstructured":"Kondrashova A, Viskari H, Haapala A-M, et al. Serological evidence of thyroid autoimmunity among schoolchildren in two different socioeconomic environments. J Clin Endocrinol Metabol. 2008;93:729\u201334.","journal-title":"J Clin Endocrinol Metabol"},{"key":"228_CR5","first-page":"281","volume":"58","author":"WR Leonard","year":"1999","unstructured":"Leonard WR, Galloway VA, Ivakine E, Osipova L, Kazakovtseva M. Nutrition, thyroid function and basal metabolism of the Evenki of central Siberia. Int J Circump Health. 1999;58:281\u201395.","journal-title":"Int J Circump Health"},{"key":"228_CR6","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1146\/annurev.anthro.34.081804.120558","volume":"34","author":"WR Leonard","year":"2005","unstructured":"Leonard WR, Snodgrass JJ, Sorensen MV. Metabolic adaptation in indigenous Siberian populations. Annu Rev Anthropol. 2005;34:451\u201371.","journal-title":"Annu Rev Anthropol"},{"key":"228_CR7","doi-asserted-by":"publisher","first-page":"814","DOI":"10.1002\/ajhb.22457","volume":"25","author":"SB Levy","year":"2013","unstructured":"Levy SB, Leonard WR, Tarskaia LA, et al. Seasonal and socioeconomic influences on thyroid function among the Yakut (Sakha) of Eastern Siberia. Am J Hum Biol. 2013;25:814\u201320.","journal-title":"Am J Hum Biol"},{"key":"228_CR8","doi-asserted-by":"publisher","first-page":"703","DOI":"10.1002\/ajhb.21200","volume":"23","author":"TJ Cepon","year":"2011","unstructured":"Cepon TJ, Snodgrass JJ, Leonard WR, et al. Circumpolar adaptation, social change, and the development of autoimmune thyroid disorders among the Yakut (Sakha) of Siberia. Am J Hum Biol. 2011;23:703\u20139.","journal-title":"Am J Hum Biol"},{"key":"228_CR9","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1038\/nrendo.2018.18","volume":"14","author":"PN Taylor","year":"2018","unstructured":"Taylor PN, Albrecht D, Scholz A, et al. Global epidemiology of hyperthyroidism and hypothyroidism. Nat Rev Endocrinol. 2018;14:301.","journal-title":"Nat Rev Endocrinol"},{"key":"228_CR10","doi-asserted-by":"publisher","first-page":"521","DOI":"10.3389\/fimmu.2017.00521","volume":"8","author":"E Fr\u00f6hlich","year":"2017","unstructured":"Fr\u00f6hlich E, Wahl R. Thyroid autoimmunity: role of anti-thyroid antibodies in thyroid and extra-thyroidal diseases. Front Immunol. 2017;8:521.","journal-title":"Front Immunol"},{"key":"228_CR11","doi-asserted-by":"publisher","first-page":"1392","DOI":"10.1093\/clinchem\/43.8.1392","volume":"43","author":"V Kaczur","year":"1997","unstructured":"Kaczur V, Vereb G, Molnar I, et al. Effect of anti-thyroid peroxidase (TPO) antibodies on TPO activity measured by chemiluminescence assay. Clin Chem. 1997;43:1392\u20136.","journal-title":"Clin Chem"},{"key":"228_CR12","doi-asserted-by":"publisher","first-page":"504","DOI":"10.1089\/thy.2015.0592","volume":"26","author":"RM Ruggeri","year":"2016","unstructured":"Ruggeri RM, Vicchio TM, Cristani M, et al. Oxidative stress and advanced glycation end products in Hashimoto\u2019s thyroiditis. Thyroid. 2016;26:504\u201311.","journal-title":"Thyroid."},{"key":"228_CR13","doi-asserted-by":"publisher","first-page":"378","DOI":"10.1093\/humupd\/dmv004","volume":"21","author":"R Vissenberg","year":"2015","unstructured":"Vissenberg R, Manders VD, Mastenbroek S, et al. Pathophysiological aspects of thyroid hormone disorders\/thyroid peroxidase autoantibodies and reproduction. Hum Reprod Update. 2015;21:378\u201387.","journal-title":"Hum Reprod Update"},{"key":"228_CR14","doi-asserted-by":"publisher","first-page":"899","DOI":"10.1353\/dem.2007.0038","volume":"44","author":"TW McDade","year":"2007","unstructured":"McDade TW, Williams S, Snodgrass JJ. What a drop can do: dried blood spots as a minimally invasive method for integrating biomarkers into population-based research. Demography. 2007;44:899\u2013925.","journal-title":"Demography."},{"key":"228_CR15","doi-asserted-by":"publisher","first-page":"307","DOI":"10.1016\/j.lab.2004.09.009","volume":"144","author":"LF Hofman","year":"2004","unstructured":"Hofman LF, Foley TP, Henry JJ, Naylor EW. The use of filter paper-dried blood spots for thyroid-antibody screening in adults. J Lab Clin Med. 2004;144:307\u201312.","journal-title":"J Lab Clin Med"},{"key":"228_CR16","doi-asserted-by":"publisher","first-page":"116","DOI":"10.1080\/19485565.2017.1283582","volume":"63","author":"GN Eick","year":"2017","unstructured":"Eick GN, Kowal P, Barrett T, Thiele EA, Snodgrass JJ. Enzyme-linked immunoassay-based quantitative measurement of apolipoprotein B (ApoB) in dried blood spots, a biomarker of cardiovascular disease risk. Biodemography Soc Biol. 2017;63:116\u201330.","journal-title":"Biodemography Soc Biol"},{"key":"228_CR17","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1136\/emj.2003.011619","volume":"22","author":"R Boyd","year":"2005","unstructured":"Boyd R, Leigh B, Stuart P. Capillary versus venous bedside blood glucose estimations. Emerg Med J. 2005;22:177\u20139.","journal-title":"Emerg Med J"},{"key":"228_CR18","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1016\/0009-8981(81)90376-4","volume":"112","author":"IR Kupke","year":"1981","unstructured":"Kupke IR, Kather B, Zeugner S. On the composition of capillary and venous blood serum. Clin Chim Acta. 1981;112:177\u201385.","journal-title":"Clin Chim Acta"},{"key":"228_CR19","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1016\/0166-0934(96)02071-X","volume":"61","author":"F Novello","year":"1996","unstructured":"Novello F, Ridolfi B, Fiore L, et al. Comparison of capillary blood versus venous blood samples in the assessment of immunity to measles. J Virol Methods. 1996;61:73\u20137.","journal-title":"J Virol Methods"},{"key":"228_CR20","doi-asserted-by":"publisher","first-page":"669","DOI":"10.1093\/cid\/civ397","volume":"61","author":"T Strecker","year":"2015","unstructured":"Strecker T, Palyi B, Ellerbrok H, et al. Field evaluation of capillary blood samples as a collection specimen for the rapid diagnosis of ebola virus infection during an outbreak emergency. Clin Infect Dis. 2015;61:669\u201375.","journal-title":"Clin Infect Dis"},{"key":"228_CR21","doi-asserted-by":"publisher","first-page":"155","DOI":"10.1046\/j.1365-2257.2001.00388.x","volume":"23","author":"ZW Yang","year":"2001","unstructured":"Yang ZW, Yang SH, Chen L, Qu J, Zhu J, Tang Z. Comparison of blood counts in venous, fingertip and arterial blood and their measurement variation. Clin Lab Haematol. 2001;23:155\u20139.","journal-title":"Clin Lab Haematol"},{"key":"228_CR22","doi-asserted-by":"publisher","first-page":"1550","DOI":"10.1016\/S0140-6736(17)30703-1","volume":"390","author":"L Chaker","year":"2017","unstructured":"Chaker L, Bianco AC, Jonklaas J, Peeters RP. Hypothyroidism. Lancet. 2017;390:1550\u201362.","journal-title":"Lancet."},{"key":"228_CR23","doi-asserted-by":"publisher","first-page":"489","DOI":"10.1210\/jcem.87.2.8182","volume":"87","author":"JG Hollowell","year":"2002","unstructured":"Hollowell JG, Staehling NW, Flanders WD, et al. Serum TSH, T4, and thyroid antibodies in the United States population (1988 to 1994): National Health and Nutrition Examination Survey (NHANES III). J Clin Endocrinol Metabol. 2002;87:489\u201399.","journal-title":"J Clin Endocrinol Metabol"},{"key":"228_CR24","doi-asserted-by":"publisher","first-page":"616","DOI":"10.1089\/thy.2016.0409","volume":"27","author":"A Amouzegar","year":"2017","unstructured":"Amouzegar A, Ghaemmaghami Z, Beigy M, et al. Natural course of euthyroidism and clues for early diagnosis of thyroid dysfunction: Tehran Thyroid Study. Thyroid. 2017;27:616\u201325.","journal-title":"Thyroid."},{"key":"228_CR25","doi-asserted-by":"publisher","first-page":"361","DOI":"10.1046\/j.1365-2265.1999.00816.x","volume":"51","author":"N Knudsen","year":"1999","unstructured":"Knudsen N, J\u00f8rgensen T, Rasmussen S, Christiansen E, Perrild H. The prevalence of thyroid dysfunction in a population with borderline iodine deficiency. Clin Endocrinol. 1999;51:361\u20137.","journal-title":"Clin Endocrinol"},{"key":"228_CR26","doi-asserted-by":"publisher","first-page":"937","DOI":"10.1089\/thy.2009.0205","volume":"19","author":"K Kasagi","year":"2009","unstructured":"Kasagi K, Takahashi N, Inoue G, Honda T, Kawachi Y, Izumi Y. Thyroid function in Japanese adults as assessed by a general health checkup system in relation with thyroid-related antibodies and other clinical parameters. Thyroid. 2009;19:937\u201344.","journal-title":"Thyroid."},{"key":"228_CR27","unstructured":"Mincer DL, Jialal I. Hashimoto thyroiditis: StatPearls; 2019."},{"key":"228_CR28","doi-asserted-by":"crossref","unstructured":"Thyroid antibodies: which, why, when and who [editorial]. Expert Rev Clin Immunol. 2006;2(5):665.","DOI":"10.1586\/1744666X.2.5.665"},{"key":"228_CR29","doi-asserted-by":"publisher","first-page":"623","DOI":"10.1055\/s-0042-112815","volume":"48","author":"M Ehlers","year":"2016","unstructured":"Ehlers M, Jordan AL, Feldkamp J, et al. Anti-thyroperoxidase antibody levels >500\u2009IU\/ml indicate a moderately increased risk for developing hypothyroidism in autoimmune thyroiditis. Horm Metab Res. 2016;48:623\u20139.","journal-title":"Horm Metab Res"}],"container-title":["Journal of Physiological Anthropology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s40101-020-00228-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s40101-020-00228-8\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s40101-020-00228-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,7,15]],"date-time":"2021-07-15T23:54:10Z","timestamp":1626393250000},"score":1,"resource":{"primary":{"URL":"https:\/\/jphysiolanthropol.biomedcentral.com\/articles\/10.1186\/s40101-020-00228-8"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,7,16]]},"references-count":29,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2020,12]]}},"alternative-id":["228"],"URL":"https:\/\/doi.org\/10.1186\/s40101-020-00228-8","relation":{},"ISSN":["1880-6805"],"issn-type":[{"value":"1880-6805","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,7,16]]},"assertion":[{"value":"20 January 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 July 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"16 July 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The Institutional Review Board approval for this study was obtained from the Committee for the Protection of Human Subjects, University of Oregon (protocol # 7062016.007), and informed consent was obtained from all participants.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Ethics approval and consent to participate"}},{"value":"All authors consent to the publication of this article.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Consent for publication"}},{"value":"None of the authors have any competing interests with regard to this manuscript to declare.","order":3,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"16"}}