Carrier-resolved technology for homogeneous and multiplexed DNA assays in a 'one-pot reaction'

The Analyst
Huan LiJianzhong Lu

Abstract

For clinical diagnosis, a small number of targets (2-10 biomarkers) are often all that is required for disease assessment and accurate early disease diagnosis. In the current paper we have developed novel, carrier-resolved, single-label-based multiplexed assays for the simultaneous detection and quantification of a limited number of DNA targets associated with breast cancer. In contrast to current encoding strategies, every hybridization signal for the corresponding DNA target in our protocol is uniquely immobilized onto one carrier vehicle with a unique and intrinsic physico-chemical signature. Moreover, a simple chemiluminescence setup is employed to read the carrier code instead of expensive and complicated flow-cytometer or imaging-systems commonly used for multiplexed assays. Herein we demonstrate a new protocol using three homogeneous carriers, i.e. thermo-sensitive poly(N-isopropylacrylamide) (PNIP), polystyrene beads, and magnetic beads respectively. This new methodology allowed for the simultaneous determination of three oligonucleotide sequences (60 bases in length) associated with the breast cancer gene (BRCA1) and showed high selectivity and attomolar-femtomolar sensitivity. The mixture of three different capture pr...Continue Reading

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Citations

Apr 5, 2011·Analytica Chimica Acta·José Antonio Murillo PulgarínIgnacio Sánchez-Ferrer Robles
May 20, 2011·Chemical Communications : Chem Comm·Zhiliang ZhangYanlin Song
May 12, 2009·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Aiping FanJianzhong Lu
Jun 18, 2015·Luminescence : the Journal of Biological and Chemical Luminescence·Hongyan CuiJianzhong Lu
Jun 18, 2021·Advanced Materials·Xinyi LiuWanwan Li

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