Screening and electrochemical detection of an antibiotic producing gene in bacteria on an integrated microchip

Analytical Methods Pub Date: 2013-09-27 DOI: 10.1039/C3AY41321G

Abstract

We demonstrate the electrochemical determination of a secondary metabolite producing gene on an integrated microfluidic chip. The fabricated microchip was assembled with a continuous channel for polymerase chain reaction (PCR) and an electrochemical detector in order to achieve rapid and sensitive determination of the valC gene. valC is a gene responsible for producing antibiotic validamycin A in Streptomyces hygroscopicus. Biotin-conjugated primers amplified the valC gene. After the PCR, a DNA amplicon was analyzed in the electrochemical cell containing a streptavidin functionalized Au working electrode. The guanosine present in the DNA amplicon released electrons upon electrochemical oxidation at 0.93 V and the peak current linearly increased with the concentration of the captured DNA amplicon. The fabricated chip successfully amplified and detected the valC gene as low as 30 pg μL?1 resulting in a sensitive, portable and integrated DNA analysis for a secondary metabolite.

Graphical abstract: Screening and electrochemical detection of an antibiotic producing gene in bacteria on an integrated microchip
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