Industry Viable Electrochemical DNA Detection Sensor Architecture via a Stem-Loop Methylene Blue Redox Reporter and Rapid in Situ Probe Immobilization Method for Pharmacogenetic Biomarker Testing Application

Asanka Jayawardena, Sher Maine Tan, Mark B. Richardson, Jianxiong Chan, Helmut Thissen, Nicolas H. Voelcker, Patrick Kwan

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Abstract

Identification of biomarkers in clinical applications for diagnostics at the point-of-care (POC) setting requires the development of industry viable biosensing platform. Herein, we report such development of biosensor architecture for the detection of pharmacogenetic biomarker HLA-B∗15:02 gene. The biosensor architecture comprises of an oligonucleotide stem-loop probe modified with a methylene blue redox (MB) reporter, immobilized via a rapid "printing"method on the commercially available disposable screen-printed electrodes (SPE). The square wave voltammetric measurements on the DNA sensor showed a clear peak difference of ∼80 nA with a significant difference in peak height values of the faradaic current generated for the MB redox moiety between the positive control (biotin-modified 19 based oligonucleotides with the sequence mimicking the specific region of the HLA-B∗15:02 allele and complementary to the probe sequence) and negative control samples (biotin-modified 19 based oligonucleotides with the sequence unrelated to the probe sequence and the HLA-B∗15:02 allele). These initial proof of concept results provide support for the possibility of using this signal-off biosensor architecture in the intended pharmacogenetic biomarker testing.

Original languageEnglish
Article number017508
Number of pages6
JournalJournal of the Electrochemical Society
Volume169
Issue number1
DOIs
Publication statusPublished - Jan 2022

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