Optimization of an enzyme linked DNA aptamer assay for cardiac troponin i detection

Synchronous multiple sample analysis on an integrated microfluidic platform

Priya Gopinathan, Anirban Sinha, Yi Da Chung, Shu-Chu Shiesh, Gwo Bin Lee

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

In this study, an enzyme linked DNA aptamer based assay was optimized for human cardiac troponin I (cTnI) detection which is a prominent biomarker for acute myocardial infarction (AMI), on an integrated microfluidic platform. This platform allowed for the multiplex detection of six samples (5 μL per sample), and only 30 min were required for detection. First, cTnI-specific aptamers were surface-coated on magnetic beads. Bead-captured proteins were allowed to bind to a primary cTnI antibody and then to a secondary antibody labelled with horseradish peroxidase. Finally, chemiluminescence intensities were detected for quantification of cTnI. Purified proteins, serum from AMI patients and unknown serum samples were used to test the efficacy of the on-chip system. The limit of detection was measured to be only 12 ng L-1, and off-target effects from other proteins were minimal. This sensitive, cTnI-specific aptamer-based assay could consequently be used for reliable diagnosis of AMI.

Original languageEnglish
Pages (from-to)4943-4951
Number of pages9
JournalAnalyst
Volume144
Issue number16
DOIs
Publication statusPublished - 2019 Aug 21

Fingerprint

Nucleotide Aptamers
Troponin
Troponin I
Microfluidics
Assays
DNA
Enzymes
assay
enzyme
Proteins
Antibodies
antibody
protein
Myocardial Infarction
serum
Chemiluminescence
Biomarkers
biomarker
Horseradish Peroxidase
Luminescence

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry
  • Biochemistry
  • Environmental Chemistry
  • Spectroscopy
  • Electrochemistry

Cite this

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abstract = "In this study, an enzyme linked DNA aptamer based assay was optimized for human cardiac troponin I (cTnI) detection which is a prominent biomarker for acute myocardial infarction (AMI), on an integrated microfluidic platform. This platform allowed for the multiplex detection of six samples (5 μL per sample), and only 30 min were required for detection. First, cTnI-specific aptamers were surface-coated on magnetic beads. Bead-captured proteins were allowed to bind to a primary cTnI antibody and then to a secondary antibody labelled with horseradish peroxidase. Finally, chemiluminescence intensities were detected for quantification of cTnI. Purified proteins, serum from AMI patients and unknown serum samples were used to test the efficacy of the on-chip system. The limit of detection was measured to be only 12 ng L-1, and off-target effects from other proteins were minimal. This sensitive, cTnI-specific aptamer-based assay could consequently be used for reliable diagnosis of AMI.",
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Optimization of an enzyme linked DNA aptamer assay for cardiac troponin i detection : Synchronous multiple sample analysis on an integrated microfluidic platform. / Gopinathan, Priya; Sinha, Anirban; Chung, Yi Da; Shiesh, Shu-Chu; Lee, Gwo Bin.

In: Analyst, Vol. 144, No. 16, 21.08.2019, p. 4943-4951.

Research output: Contribution to journalArticle

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