G-quadruplex-based assay combined with aptamer and gold nanoparticles for Escherichia coli K88 determination

A colorimetric method was developed using G-quadruplex and gold nanoparticles (AuNPs) for determination of Escherichia coli K88 (ETEC K88). It was composed of two modules: (1) an aptamer as biorecognizing element and (2) a capturing DNA (modified with AuNPs at 5′) as a transducer. In the absence of target bacteria, the aptamer can form stable double strands with capturing DNA, preventing the binding of capturing DNA to the G-quadruplex. However, the double strands of capturing DNA and aptamer are untied due to the stronger binding of aptamers to bacteria in the presence of target bacteria. As a result, the G-quadruplex binds to capture DNA and leads to the aggregation and color change of AuNPs, which can be monitored by a spectrophotometer or visualization. The quantitative determination was achieved by monitoring the optical density change of AuNPs solution at 524 nm after target addition. Under optimal conditions, the method has a low detection limit (1.35 × 102 CFU mL−1) and a linear response in the range 102 to 106 CFU mL−1.