Triple-helix molecular switch-based versatile 'off-on' electrochemiluminescence and fluorescence biosensing platform for ultrasensitive detection of lipopolysaccharide by multiple-amplification strategy

摘要

Herein, a novel biosensing platform for versatile electrochemiluminescence (ECL) "off" and fluorescence (FL) "on" detection of lipopolysaccharide (LPS) with multiple-amplification strategy is proposed. The specific recognition of target to aptamer on the magnetic beads (MB) firstly released abundant DNA sequences of three kinds. The sequences hybridized with multifunctional molecular beacon (MMB) and initiated numerous bidirectional polymerization and shearing reactions, generating a large number of DNA fragments (a1) by multiple cycling amplification. Then a1 was introduced to the triple-helix sensing system, opening the triple-helix structure. In ECL system, the G-rich chains S2 were exposed to form G-quadruplex-hemin complex in the presence of hemin, which could efficiently quench ECL for "off" detection of LPS. In FL system, the fluorophore FAM and quencher BHQ on S1 chain were separated with opening of triple-helix structure, achieving fluorescence "on" signal for LPS assay. So the versatile platform can achieve greatly amplified ECL and FL signal changes for sensitive assay of LPS, showing wide linear ranges (0.1 fg/mL-0.1 ng/mL by ECL and 10 fg/mL(-1)-1 mu g/mL by FL) and low detection limits (0.012 fg/mL by ECL and 1.269 fg/mL by FL). Therefore, the present ECL "Off" and FL "On" dual-signal detection patterns for LPS displayed many advantages over other reported methods, which provided an outlook for future applications in clinical diagnosis.