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首页> 外文期刊>Journal of Physiological Anthropology >Development and validation of an ELISA for a biomarker of thyroid dysfunction, thyroid peroxidase autoantibodies (TPO-Ab), in dried blood spots
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Development and validation of an ELISA for a biomarker of thyroid dysfunction, thyroid peroxidase autoantibodies (TPO-Ab), in dried blood spots

机译：甲状腺功能障碍生物标志物的开发和验证甲状腺功能障碍，甲状腺过氧化物酶自身抗体（TPO-AB），干燥血斑

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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.
机译：背景：在过去的世纪过去，过敏性和自身免疫条件的患病率一直在稳步增加。一个假设提出来解释这是旧的朋友假设，它的假设增加了卫生，城市化和生活方式的变化，这使我们暴露于我们共同进化的寄生虫和微生物，导致免疫失调。然而，在传统的生活中的研究，暴露于更大的寄生虫和病原体载荷，例如在我们的进化期间遇到的那些，部分是由于缺乏自身免疫疾病的微创，现场友好的生物标志物。因此，我们开发了一种ELISA，用于评估甲状腺过氧化物酶的阳性阳性菌（TPO-AB），基于干血斑（DBS）样品的自身免疫性甲状腺疾病的指标。结果：我们使用Accubind抗甲状腺过氧化物酶测试系统来筛选我们的验证样品包括来自182名成人的匹配的Fingerprick DBS，静脉DB和血浆样品。在确认我们在我们的验证样本中有TPO-AB阳性的个体（n = 12），我们开发了一个间接的ELISA，以测量从一个3毫米DBS打孔的TPO-AB水平。 DBS样品的测定敏感性和特异性分别使用≥26Ω·IU / mL的截止值，分别为91.7-100％和98.2-98.8％。测定重复质量控制DBS冲头的测定可靠性为5.2％，而测定的可靠性范围为高，培养基和低DBS控制的11.5-24.4％。稀释线性范围为80至120％，尖峰和恢复实验表明，DBS基质不会干扰TPO-AB的检测。 DBS样品中的TPO-AB水平仍然是稳定的 - 28？°C或-80？°C，但随着时间的推移，在22Ω·℃或37℃的DBS样品中随着时间的推移而降低：我们开发了一个 - 房屋，套件独立的间接ELISA测定以确定基于干血斑的个体TPO-AB阳性，代表一种具有在一系列研究环境中的潜在应用的成本效益的方法。

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