Agglutination-PCR

Agglutination-PCR (ADAP) is an ultrasensitive solution-phase method for detecting antibodies.[1] Antibodies bind to and agglutinate synthetic antigen–DNA conjugates, enabling ligation of the DNA strands and subsequent quantification by qPCR. Like other Immuno-PCR (IPCR) detection methods[2][3] ADAP combines the specificity of antibody-antigen recognition and the sensitivity of PCR. ADAP detects zepto- to attomoles of antibodies in 2 μL of sample with a dynamic range spanning 5–6 orders of magnitude. For example, ADAP allows to detect anti-thyroglobulin autoantibodies from human patient plasma with a 1000-fold increased sensitivity over an FDA-approved radioimmunoassay. ADAP also allows to simultaneously detect multiple antibodies in one experiment, much more than ELISA or radioimmunoassay.

The study published in the ACS Central Science journal mentioned that this testing method is 10,000 times more effective than the existing diagnostic techniques.[4][5]

Another advantage of ADAP method is the simplicity. It can be adapted to such a cheap equipment as, for example, SlipChip.[6][7] It does not require complex and expensive equipment, it does not require working with hazardous radioactive reagents.

References
  1. Tsai, Cheng-Ting; Robinson, Peter V.; Spencer, Carole A.; Bertozzi, Carolyn R. (2016). "Ultrasensitive Antibody Detection by Agglutination-PCR (ADAP)". ACS Central Science. 2 (3): 139–147. doi:10.1021/acscentsci.5b00340. PMC 4819452. PMID 27064772.
  2. Chang, Le; Li, Jinming; Wang, Lunan (2016). "Immuno-PCR: An ultrasensitive immunoassay for biomolecular detection". Analytica Chimica Acta. 910: 12–24. doi:10.1016/j.aca.2015.12.039. PMID 26873464.
  3. Spengler, Mark; Adler, Michael; Niemeyer, Christof M. (2015). "Highly sensitive ligand-binding assays in pre-clinical and clinical applications: Immuno-PCR and other emerging techniques". The Analyst. 140 (18): 6175–6195. Bibcode:2015Ana...140.6175S. doi:10.1039/C5AN00822K. PMID 26196036.
  4. University, Stanford (2016-03-10). "Stanford chemists develop an ultra-sensitive test for cancers, HIV". Stanford News. Retrieved 2017-09-12.
  5. "New Ultra-Sensitive Test Up to 10,000 Times More Effective In Detecting HIV, Cancer". Tech Times. 2016-03-21. Retrieved 2017-09-12.
  6. Shen, Feng; Du, Wenbin; Kreutz, Jason E.; Fok, Alice; Ismagilov, Rustem F. (2010). "Digital PCR on a Slip Chip". Lab on a Chip. 10 (20): 2666–2672. doi:10.1039/c004521g. PMC 2948063. PMID 20596567.
  7. Rodriguez-Manzano, Jesus; Karymov, Mikhail A.; Begolo, Stefano; Selck, David A.; Zhukov, Dmitriy V.; Jue, Erik; Ismagilov, Rustem F. (2016). "Reading Out Single-Molecule Digital RNA and DNA Isothermal Amplification in Nanoliter Volumes with Unmodified Camera Phones". ACS Nano. 10 (3): 3102–3113. doi:10.1021/acsnano.5b07338. PMC 4819493. PMID 26900709.

Further reading
  • Alexander, L., Rasmus, B., Charlotte, B., & Daniel, A. (2023). Antibody detection by agglutination-PCR (ADAP) assays for the analysis of tissue transglutaminase autoantibodies in celiac disease. Journal of Immunological Methods, 113502. PMID 37257686 doi:10.1016/j.jim.2023.113502
  • Lind, A., de Jesus Cortez, F., Ramelius, A., Bennet, R., Robinson, P. V., Seftel, D., ... & Lernmark, Å. (2022). Multiplex agglutination-PCR (ADAP) autoantibody assays compared to radiobinding autoantibodies in type 1 diabetes and celiac disease. Journal of Immunological Methods, 506, 113265. PMID 35358496 doi:10.1016/j.jim.2022.113265
  • de Jesus Cortez, F., Gebhart, D., Tandel, D., Robinson, P. V., Seftel, D., Wilson, D. M., ... & Tsai, C. T. (2022). Automation of a multiplex agglutination-PCR (ADAP) type 1 diabetes (T1D) assay for the rapid analysis of islet autoantibodies. SLAS technology, 27(1), 26-31. PMID 35058202 doi:10.1016/j.slast.2021.10.001
This article is issued from Wikipedia. The text is licensed under Creative Commons - Attribution - Sharealike. Additional terms may apply for the media files.