Integrated Microfluidic Device With Carbon-Thread Microelectrodes for Electrochemical DNA Elemental Analysis

Evidently, any alternation in the concentration of the essential DNA elements, adenine (A), guanine (G), cytosine (C), and thymine (T), leads to several deformities in the physiological process causing various disorders. So, to realize a simple and precise technique for simultaneous determination of the DNA elements continue to remain a challenge. Microfluidic devices offer numerous advantage, such as low volume consumption, rapid response, highly sensitive and accurate real time analysis, for point of care testing (POCT). Herein, a microfluidic electrochemical device has been developed with three electrodes fabricated using a carbon-thread microelectrode (CTME) for DNA elemental detection. CTME, functionalized with graphitize mesoporous carbon (GMC), worked as a working electrode, bare CTME functioned as an auxiliary electrode while CTME coated with Ag/AgCl ink performed as a reference electrode. The developed device was used for evaluating individual DNA elemental base pairs simultaneously using various electrochemical techniques. The anodic peak current obtained for the DNA bases were 0.56 ± 0.04 V (G), 0.92 ± 0.02 V (A), 1.09 ± 0.05 V (T) and 1.24 ± 0.04 V (C) in a potential window of 0.2 V to 1.5 V. The device was corroborated for simultaneous sensing, and detection limits were found to be
$36.73 ~\mu \text{M}$
(G),
$20 ~\mu \text{M}$
(A),
$22 ~\mu \text{M}$
(T) and
$19.78 ~\mu \text{M}$
(C) in the linear range of
$50 ~\mu \text{M}$
–
$500 ~\mu \text{M}$
. Finally, the device was successfully used for instantaneous determination of DNA bases in the human blood serum sample. Overall, this work demonstrates the use of a simple microfluidic device with CTMEs for electrochemical determination of DNA bases amenable for POCT.