Development of a genosensor for peanut allergen ARA h 2 detection and its optimization by surface response...

Area of Science:

• Biosensors
• Analytical Chemistry
• Food Allergen Detection

Background:

• Peanut allergy is a significant health concern, often triggered by the allergen Ara h 2.
• Accurate detection of food allergens is crucial for consumer safety and regulatory compliance.

Purpose of the Study:

• To develop a selective electrochemical genosensor for detecting a specific DNA sequence of the peanut allergen Ara h 2.
• To optimize the genosensor's performance using a Design of Experiments (DoE) approach.

Main Methods:

• Fabrication of screen-printed gold electrodes with immobilized oligonucleotides via S-Au bonds.
• Utilized mixed self-assembled monolayers (SAMs) of capture probes and mercaptohexanol to prevent non-specific adsorption.
• Employed a sandwich assay format to enhance detection efficiency and avoid target labeling.

Main Results:

• Optimized capture probe concentration at 1.34 µM and mercaptohexanol concentration at 3.15 mM.
• Achieved a high sensitivity of 3 µA/nM.
• Demonstrated a linear detection range from 5×10⁻¹¹ to 5×10⁻⁸ M with a detection limit of 10 pM.

Conclusions:

• The developed genosensor provides a sensitive and selective method for detecting Ara h 2 DNA.
• The optimized electrochemical approach offers a promising tool for rapid and reliable allergen monitoring in food products.