Label-Free Liquid Crystal Aptamer Sensors Based on Single-Stranded Nucleic Acid π-Structures for Detecting cTnI

Cardiac troponin I (cTnI) is a highly sensitive and important serological marker for clinical diagnosis of myocardial injury. Its rapid detection is crucial for the early diagnosis of cardiovascular diseases such as acute myocardial infarction. In this study, based on nucleic acid molecular hybridization and aptamer-specific binding to target molecules, a label-free liquid crystal aptamer sensor based on single-stranded nucleic acid π-structures was developed and applied for the quantitative detection of cTnI. The CP1 and CP2 oligonucleotide chains, complementary to the bases at both ends of the aptamer, are covalently bonded to the sensor substrate via APTES and GA-mediated molecules. The aptamer forms a π-structure with CP1 and CP2 through nucleic acid hybridization, serving as a target molecule capture probe. When cTnI is present in the system, cTnI and the complementary oligonucleotide chains competitively bind with the aptamer, causing the breakdown of the π-structure within the sensor. This reinstates the long-range ordered alignment of the 5CB liquid crystal molecules within the sensor, enabling quantitative measurement of cTnI through variations in optical images. Experimental results show that within the range of 0.01 to 25 ng/mL for cTnI concentration, there is a linear correlation between the brightness area coverage (Br) in the polarized light microscopy images of the sensor and the logarithm of the cTnI concentration, with a correlation coefficient (r). The detection limit is 5.16 pg/mL. This method is label-free, simple to operate, and low-cost, with good specificity and a low detection limit, achieving cTnI detection in serum samples.