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Automation of a multiplex agglutination-PCR (ADAP) type 1 diabetes (T1D) assay for the rapid analysis of islet autoantibodies
SLAS Technology: Translating Life Sciences Innovation ( IF 2.5 ) Pub Date : 2021-10-25 , DOI: 10.1016/j.slast.2021.10.001
Felipe de Jesus Cortez 1 , David Gebhart 1 , Devangkumar Tandel 1 , Peter V. Robinson 1 , David Seftel 1 , Darrell M. Wilson 2, 3 , David M. Maahs 2, 3 , Bruce A. Buckingham 2, 3 , Kevin W.P. Miller 4 , Cheng-ting Tsai 1
Affiliation  

Screening for islet autoantibody markers to identify individuals who are at high risk for developing type 1 diabetes (T1D), often years in advance of clinical symptoms, is both a challenge and a necessity. Identifying high-risk individuals not only reduces hospitalization and rates of life-threatening diabetes ketoacidosis (DKA), but also directs enrollment into prevention trials that require patients who are in the early stages of disease. Here we describe an automated high-throughput multiplex islet autoantibody assay that integrates antibody detection by agglutination-PCR (ADAP) chemistry on the Hamilton Microlab STAR liquid handling platform. The automated system features on-deck thermal cycling and plate sealing to minimize the level of human intervention. The automated multiplex ADAP T1D assay performed similarly to that of manual methods using two distinct cohorts of clinical specimens obtained from the Lucile Packard Children's Hospital at Stanford University and the 2018 Islet Autoantibody Standardization Program (IASP). Notably, the automated assay requires only 4 μL of serum sample for the simultaneous analysis of GAD, IA-2 and insulin autoantibodies.

Up to 96 samples may be processed in as little as 3 hours, and the only user intervention required is to transfer a final sealed 96-well plate containing PCR amplicons onto a quantitative PCR (RT-qPCR) instrument for quantification. The automated system is particularly well suited for large-scale analysis of islet autoantibodies in a reproducible, timely, and cost-effective manner.



中文翻译:

用于快速分析胰岛自身抗体的多重凝集 PCR (ADAP) 1 型糖尿病 (T1D) 检测的自动化

筛查胰岛自身抗体标志物以识别具有 1 型糖尿病 (T1D) 高风险的个体,通常比临床症状提前数年,既是挑战也是必要的。识别高风险个体不仅可以降低住院率和危及生命的糖尿病酮症酸中毒 (DKA) 的发生率,而且还可以指导患者参与需要处于疾病早期阶段的患者的预防试验。在这里,我们描述了一种自动化的高通量多重胰岛自身抗体检测,该检测在 Hamilton Microlab STAR 液体处理平台上集成了通过凝集 PCR (ADAP) 化学进行的抗体检测。该自动化系统具有甲板热循环和板密封功能,可最大程度地减少人为干预。自动多重 ADAP T1D 测定的执行类似于手动方法,使用从斯坦福大学 Lucile Packard 儿童医院和 2018 年胰岛自身抗体标准化计划 (IASP) 获得的两个不同的临床标本队列。值得注意的是,自动化检测仅需 4 μL 血清样本即可同时分析 GAD、IA-2 和胰岛素自身抗体。

最多可在 3 小时内处理多达 96 个样本,唯一需要的用户干预是将包含 PCR 扩增子的最终密封 96 孔板转移到定量 PCR (RT-qPCR) 仪器上进行定量。该自动化系统特别适合以可重复、及时和经济高效的方式对胰岛自身抗体进行大规模分析。

更新日期:2021-10-25
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