Microfluidic device using chemiluminescence and a DNA-arrayed thin film transistor photosensor for single nucleotide polymorphism genotyping of PCR amplicons from whole blood

Keiichi Hatakeyama, Tsuyoshi Tanaka, Masahiro Sawaguchi, Akihito Iwadate, Yasushi Mizutani, Kazuhiro Sasaki, Naofumi Tateishi, Tadashi Matsunaga

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

This work describes a novel microfluidic device using a thin film transistor (TFT) photosensor integrating a microfluidic channel, a DNA chip platform, and a photodetector for the discrimination of single nucleotide polymorphisms (SNPs). A DNA-arrayed TFT photosensor was used as a DNA chip platform and photo detecting device. Chemiluminescence was used for DNA sensing because chemiluminescence provides higher sensitivity and requires simpler instrumentation than fluorescence methods. The SNP of biotinylated target DNA was detected based on chemiluminescence by using horse radish peroxidase-conjugated streptavidin. The lower detection limit for a model biotinylated oligonucleotide (63-mer) was 0.5 nM, much lower than expected DNA concentrations in a practical application of this device. Furthermore, SNP detection in the aldehyde dehydrogenase 2 gene was successfully achieved using DNA-arrayed TFT photosensor without DNA extraction and DNA purification using PCR products. The assay was completed in less than one hour. Our technology will be a promising approach to developing a miniaturized, disposable DNA chip with high sensitivity.

Original languageEnglish
Pages (from-to)1052-1058
Number of pages7
JournalLab on a Chip
Volume9
Issue number8
DOIs
Publication statusPublished - 2009
Externally publishedYes

Fingerprint

Lab-On-A-Chip Devices
Chemiluminescence
Thin film transistors
Nucleotides
Luminescence
Polymorphism
Microfluidics
Single Nucleotide Polymorphism
DNA
Blood
Polymerase Chain Reaction
Oligonucleotide Array Sequence Analysis
Raphanus
Equipment and Supplies
Aldehyde Dehydrogenase
Streptavidin
Oligonucleotides
Peroxidase
Limit of Detection
Fluorescence

ASJC Scopus subject areas

  • Bioengineering
  • Chemistry(all)
  • Biochemistry
  • Biomedical Engineering

Cite this

Microfluidic device using chemiluminescence and a DNA-arrayed thin film transistor photosensor for single nucleotide polymorphism genotyping of PCR amplicons from whole blood. / Hatakeyama, Keiichi; Tanaka, Tsuyoshi; Sawaguchi, Masahiro; Iwadate, Akihito; Mizutani, Yasushi; Sasaki, Kazuhiro; Tateishi, Naofumi; Matsunaga, Tadashi.

In: Lab on a Chip, Vol. 9, No. 8, 2009, p. 1052-1058.

Research output: Contribution to journalArticle

Hatakeyama, Keiichi ; Tanaka, Tsuyoshi ; Sawaguchi, Masahiro ; Iwadate, Akihito ; Mizutani, Yasushi ; Sasaki, Kazuhiro ; Tateishi, Naofumi ; Matsunaga, Tadashi. / Microfluidic device using chemiluminescence and a DNA-arrayed thin film transistor photosensor for single nucleotide polymorphism genotyping of PCR amplicons from whole blood. In: Lab on a Chip. 2009 ; Vol. 9, No. 8. pp. 1052-1058.
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