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

Advances in risk predictive performance of pre-symptomatic type 1 diabetes via the multiplex Antibody-Detection-by-Agglutination-PCR assay

“…Previously, we reported a multiplex ADAP method for detecting three islet autoantibodies ( 15 ). In addition, we described an automated Hamilton MicroLab STAR system to carry out the 3-plex ADAP assay ( 16 ). Recently, we expanded the assay to 5-plex to test for IAA, GADA, IA2A, ZnT8A, and TGA on a modified version of Hamilton MicroLab STAR to achieve full automation ( 17 ).…”

Assay methods based on proximity-enhanced reactions for detecting non-nucleic acid molecules

“…Proteins based on alternative scaffolds—e.g., designed ankyrin repeat proteins (DARPins)—have also been developed as binders for targets and have been used to generate binder–oligonucleotide conjugates ( Gu et al, 2013 ) ( Table 1 ). Additionally, antigens have been used as binders for the detection of antibodies ( Tsai et al, 2016 ; Tsai et al, 2018 ; Cortez et al, 2020 ; Karp et al, 2020 ; Cortez et al, 2022 ; Lind et al, 2022 ) ( Table 1 ). Aptamers have also been used as binders ( Fredriksson et al, 2002 ; Yang and Ellington, 2008 ; Joonyul Kim, 2010 ; Liu et al, 2020 ; Zhao et al, 2020 ; Marnissi et al, 2021 ) ( Table 1 ); they possess a key advantage over protein binders in that binder (aptamer)–oligonucleotide molecules can be prepared as a single molecule via chemical or biological pathways ( Fredriksson et al, 2002 ; Yang and Ellington, 2008 ; Joonyul Kim, 2010 ; Liu et al, 2020 ; Zhao et al, 2020 ; Marnissi et al, 2021 ).…”

“…Methods of monitoring the amplification process in a real-time manner, usually via tracking fluorescent signals, are known as quantitative PCR (qPCR). They can quantify the abundance of template DNA with very high sensitivity and specificity ( Fredriksson et al, 2002 ; Gullberg et al, 2003 ; Gullberg et al, 2004 ; Gustafsdottir et al, 2006 ; Fredriksson et al, 2007 ; Schallmeiner et al, 2007 ; Fredriksson et al, 2008 ; Yang and Ellington, 2008 ; Chang et al, 2009 ; Joonyul Kim, 2010 ; Masood Kamali-Moghaddam and Gholamreza Tavoosidana, 2010 ; Lundberg et al, 2011b ; Tavoosidana et al, 2011 ; Cheng et al, 2012 ; Gu et al, 2013 ; Blokzijl et al, 2014 ; Dhillon et al, 2016 ; Robinson et al, 2016 ; Tsai et al, 2016 ; Jalili et al, 2018 ; Tsai et al, 2018 ; Li et al, 2019 ; Cortez et al, 2020 ; Karp et al, 2020 ; Liu et al, 2020 ; Marnissi et al, 2021 ; Song et al, 2021 ; Chen and Liang, 2022 ; Cortez et al, 2022 ; Lind et al, 2022 ) ( Table 1 ). Recently, next-generation sequencing (NGS) following a short PCR has been used to detect PLA products ( Darmanis et al, 2011 ; Nong et al, 2013 ).…”

“…The first PLA method was developed to detect platelet-derived growth factor using two aptamer binders and qPCR ( Fredriksson et al, 2002 ). Since the development of this method, PLA methods coupled with PCR have been developed to detect various non-nucleic acid target molecules, including cytokines ( Gullberg et al, 2004 ), pathogens ( Gustafsdottir et al, 2006 ; Wang et al, 2021 ), thrombin ( Yang and Ellington, 2008 ; Joonyul Kim, 2010 ), clenbuterol ( Cheng et al, 2012 ), ractopamine ( Cheng et al, 2012 ), toxins ( Dhillon et al, 2016 ), antibodies ( Tsai et al, 2016 ; Tsai et al, 2018 ; Cortez et al, 2020 ; Karp et al, 2020 ; Cortez et al, 2022 ), viruses ( Liu et al, 2020 ; Marnissi et al, 2021 ), and post-translationally modified proteins ( Robinson et al, 2016 ; Oliveira et al, 2018 ; Song et al, 2021 ; Chen and Liang, 2022 ) ( Table 1 ). Multiplexed protein marker detection using PLA methods has been reported using qPCR ( Fredriksson et al, 2007 ; Fredriksson et al, 2008 ; Chang et al, 2009 ; Lundberg et al, 2011b ; Blokzijl et al, 2014 ) ( Table 1 ).…”

Childhood screening for type 1 diabetes comparing automated multiplex Antibody Detection by Agglutination-PCR (ADAP) with single plex islet autoantibody radiobinding assays

“… 24 Prior attempts to replace the RBA with multiplex autoantibody assays include two-sided ELISA for GADA, IA-2A and ZnT8A, 25 , 26 luciferase immunoprecipitation system (LIPS), 27 multiplex assay by electrochemiluminescence (ECL) 28 and the Antibody Detection by Agglutination-PCR (ADAP) assay. 29 , 30 , 31 , 32 …”