Fast Aptamer Generation Method Based on the Electrodynamic Microfluidic Channel and Evaluation of Aptamer Sensor...

Area of Science:

• Biotechnology
• Analytical Chemistry
• Biomolecular Engineering

Background:

• Developing rapid and sensitive diagnostic tools for malaria is crucial for disease control.
• Existing aptamer selection methods can be time-consuming and complex.
• Plasmodium vivax lactate dehydrogenase (PvLDH) is a key biomarker for malaria diagnosis.

Purpose of the Study:

• To establish a fast aptamer generation method using a screen-printed electrodynamic microfluidic channel device.
• To develop a sensitive biosensor for detecting the malaria biomarker PvLDH.
• To demonstrate the efficiency of AC electric fields in aptamer selection.

Main Methods:

• A screen-printed electrodynamic microfluidic device was utilized for aptamer selection.
• Plasmodium vivax lactate dehydrogenase (PvLDH) was immobilized on channel walls.
• An AC electric field was applied to induce specific binding between PvLDH and DNA library molecules.
• Quantitative polymerase chain reaction (qPCR) was used to confirm partitioning efficiency.
• Selected aptamers were isolated, sequenced, and characterized using electrochemical impedance spectroscopy (EIS) and fluorescence methods.

Main Results:

• A high partitioning efficiency of 1.67 × 10^7 was achieved for PvLDH and DNA library binding.
• Five distinct aptamers were generated in a single selection cycle.
• The dissociation constants (Kd) of the selected aptamers were determined.
• The most sensitive aptamer exhibited a detection limit of 7.8 ± 0.4 fM for PvLDH.
• The developed sensor demonstrated high reliability compared to other malaria sensors.

Conclusions:

• The screen-printed electrodynamic microfluidic device offers a fast and efficient platform for aptamer generation.
• The selected aptamers are highly specific and sensitive for PvLDH detection.
• This technology holds promise for the development of rapid and reliable malaria diagnostic tools.