Authors:
Tanu Bhardwaj
1
and
Sandeep Kumar Jha
2
Affiliations:
1
Indian Institute of Technology, India
;
2
Indian Institute of Technology Delhi, India
Keyword(s):
PMMA, Microfluidics, Aptamer, Biosensing, FAM, Fluorescence.
Abstract:
In this work, we are reporting fabrication of a simple and low cost setup for fluorescence detection based on aptamer probes. For this reason, we fabricated a PMMA-PMMA microfluidic chip using easily available laboratory techniques and combined the chip with a simple fluorescence detection setup using optical fiber, filter, detector and a commercial spectroscopy software. In this new approach, we used two different strategies to use aptamers as probe. In first strategy, detection of any nucleic acid could be targeted using simple DNA hybridization with aptamer probe. Such strategy can be used in analysis of samples with specific nucleic acid sequence, such as pathogen. We proved this using known sequence of ssDNA aptamer probe immobilized on detection zone on microchip and its FAM labeled complementary strand was passed over it using microfluidic condition. In other strategy, we attempted detection of any protein or biomarker using sandwich fluorimetric technique with primary and lab
eled secondary aptamer immobilized on sensing region. For this, we used thrombin as model target to validate our setup. Both the strategies proved satisfactory on our setup. Even more, LOD was also impressive. In future, this setup could further be miniaturized by using a small on-chip CCD array detector, microcontroller based electronics and LabVIEW software based control.
(More)