Barbara Davis Center for Diabetes, University of Colorado School of Medicine.
Barbara Davis Center for Diabetes, University of Colorado School of Medicine; Department of Endocrinology, Guangzhou First People's Hospital, School of Medicine, South China University of Technology.
J Vis Exp. 2022 Jul 5(185). doi: 10.3791/63787.
An ongoing clinical trial, Autoimmunity Screening for Kids (ASK), is the first screening study in the general population for type 1 diabetes (T1D) and celiac disease in the United States. With the coronavirus disease 2019 (COVID-19) pandemic, the epidemiology of COVID-19 in the general population and knowledge about the association between COVID-19 infection and T1D development are urgently needed. The currently standard screening method of the radio-binding assay (RBA) has met two great challenges: low efficiency with a single assay format and low disease specificity with a large proportion of low-affinity antibodies generated in screening. With the platform of the multiplex electrochemiluminescence (ECL) assay we established previously, a novel 6-Plex ECL assay was developed that combines, in a single well, all four islet autoantibodies (IAbs) to insulin, glutamic acid decarboxylase (GAD65), insulinoma antigen 2 (IA-2), and Zinc transporter 8 (ZnT8) for T1D, transglutaminase autoantibodies (TGA) for celiac disease, and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor-binding domain (RBD) antibodies for COVID-19. The assay was validated in blind using 880 samples from the ASK study, including 325 positive samples and 555 all antibody-negative samples, and compared with the standard RBAs and a single ECL assay. With the advantages of high efficiency, low cost, and low serum volume, this assay has been accepted as the primary screening tool for the ASK study.
一项正在进行的临床试验——儿童自身免疫筛查(ASK),是美国首例针对 1 型糖尿病(T1D)和乳糜泻的一般人群筛查研究。由于 2019 年冠状病毒病(COVID-19)大流行,人们迫切需要了解 COVID-19 在一般人群中的流行病学情况,以及 COVID-19 感染与 T1D 发展之间的关联。目前,放射结合测定(RBA)作为标准的筛查方法,面临着两个巨大的挑战:单一检测方法效率低下,以及筛查中产生的大量低亲和力抗体导致疾病特异性低。我们先前建立了基于多重电化学发光(ECL)检测的平台,开发了一种新的 6-Plex ECL 检测方法,可在单个孔中同时结合胰岛素、谷氨酸脱羧酶(GAD65)、胰岛抗原 2(IA-2)和锌转运蛋白 8(ZnT8)这 4 种胰岛自身抗体(IAbs)用于 T1D 的检测、针对乳糜泻的转谷氨酰胺酶自身抗体(TGA)检测,以及针对 COVID-19 的严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)受体结合域(RBD)抗体检测。该检测方法在 ASK 研究的 880 份盲样中进行了验证,包括 325 份阳性样本和 555 份所有抗体阴性样本,并与标准 RBA 和单一 ECL 检测进行了比较。由于具有高效、低成本和低血清用量的优势,该检测方法已被接受为 ASK 研究的主要筛查工具。