Microfluidic biosensor for the detection of DNA by fluorescence enhancement and the following streptavidin detection by fluorescence quenching
Artikel i vetenskaplig tidskrift, 2014

We reported an optical DNA/protein microfluidic sensor which consists of single stranded (ss) DNA-Cy3 probes on gold surface and simple line-shape microfluidic channel. These ssDNA-Cy3 probes with random sequence in bulk solution or on gold surface exhibits fluorescence enhancement after binding with complementary ssDNA (cssDNA) targets. Particularly it did not require complicated design or hairpin-like stem-loop conformation, which made it easier to be made and applied in analytes detection by fluorescence switching techniques. Using ssDNA-cy3 probes attached on gold surface in a microfluidic channel, strong fluorescence enhancement was measured by ssDNA with cssDNA binding or ssDNA with cssDNA-biotin binding. The following introduction of streptavidin resulted in fluorescence quenching (fluorescence decrease) because of the binding of hybridized DNA-biotin with streptavidin. This sensor showed strong affinity and high sensitivity toward the streptavidin, the minimum detectable concentration for streptavidin was 1 pM, equating to an absolute detection limit of 60 amol in this microfluidic channel. Microfluidic channel height and flow rate is optimized to increase surface reaction efficiency and fluorescence switching efficiency. In contrast to previously reported optical molecular beacon approach, this sensor can be used not only for the detection of cssDNA target, but also for the detection of streptavidin. This microfluidic sensor offers the promise of analyzing kinds of molecular targets or immunoreactions.

probes

binding proteins

interrogation

hybridization

analysis systems

sensitive detection

nucleic-acid

thiazole orange

molecular-beacon

electron-transfer

Författare

Jun Wang

Chalmers, Kemi- och bioteknik, Analytisk kemi

Michihiko Aki

Fujitsu FTP Corporation

Daisuke Onoshima

Nagoya University

Kenji Arinaga

Fujitsu FTP Corporation

Noritada Kaji

Nagoya University

Fujitsu FTP Corporation

Manabu Tokeshi

Hokkaido University

Nagoya University

Shozo Fujita

Fujitsu FTP Corporation

Naoki Yokoyama

Fujitsu FTP Corporation

Yoshinobu Baba

Nagoya University

National Institute of Advanced Industrial Science and Technology

Biosensors and Bioelectronics

0956-5663 (ISSN)

Vol. 51 280-285

Ämneskategorier

Biofysik

DOI

10.1016/j.bios.2013.07.058