Visual detection of thrombin using a strip biosensor through aptamer-cleavage reaction with enzyme catalytic amplification.

ABSTRACT

A new class of strip biosensors has been established based on well-distributed thrombin aptamer-linked gold nanoparticle aggregates, which will undergo a cracking reaction when the target recognizes its homologous aptamer. Combining the aptamer-cleavage reaction with the enzyme catalytic amplification system, our proposed lateral flow strip biosensor (LFB) is capable of visually detecting 6.4 pM of thrombin without instrumentation within 12 minutes. Under the optimal conditions, the quantitative detection of thrombin by a portable strip reader exhibited a linear relationship between the peak area and the concentration of thrombin in the range of 6.4 pM-500 nM with a detection limit of 4.9 pM, which is three orders of magnitude lower than that of the aptamer-functionalized gold nanoparticle-based LFB (2.5 nM, Xu et al., Anal. Chem., 2009, 81, 669-675). As the aptamers have no special requirements and the gold nanoparticles can also be replaced by other metallic nanoparticles, this method for strip sensing is expected to be generally applicable in point of care testing, home testing, medical diagnostics, clinical diagnosis, and environmental monitoring.