System and Method for Immobilization free electrochemiluminescence DNA detection using a luminophore dye for multi-species detection

摘要

Present invention disclosed a novel method for detecting and quantifying target DNA from the biological sample. It provides a method of amplification of DNA sequences with loop-mediated isothermal amplification (LAMP) and its easy and accurate detection and quantification by electrochemiluminescence (ECL) technique. The target LAMP DNA is bound electrostatically with [Ru(bpy)₃]⁺² on the carbon electrode surface, and an ECL reaction is triggered by tripropylamine (TPrA) to yield luminescence. This is a highly sensitive strategy for the detection of sequence-specific DNA from different biological samples at picogram levels. The target DNA of Sus scrofa (pork) meat was detected as low as 1 pg/μL (3.43×10⁻¹ copies/μL) and for Bacillus subtilis DNA samples the detection limit of 10 pg/μL (2.2×10³ copies/μL) was achieved.