Automation of a multiplex agglutination-PCR (ADAP) type 1 diabetes (T1D) assay for the rapid analysis of islet autoantibodies

SLAS Technology

Kevin W.P. Miller¹  David M. Maahs¹  Darrell M. Wilson²  Bruce A. Buckingham²  Cheng-ting Tsai²  Peter V. Robinson³  David Gebhart³  David Seftel³  Felipe de Jesus Cortez³  Devangkumar Tandel³ 

[1] Stanford Diabetes Research Center, Stanford, CA, USA;Department of Pediatrics, Division of Pediatric Endocrinology and Diabetes, Stanford, University School of Medicine, Stanford, CA, USA;Enable Biosciences Inc., South San Francisco, CA, USA;

关键词: PCR;    Antibody;    Diabetes;    Liquid handling;    Clinical automation;   

DOI  :  

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown