One-step detection for two serological biomarker species to improve the diagnostic accuracy of hepatocellular carcinoma

Abstract

At present, the accuracy of clinical hepatocellular carcinoma (HCC) diagnosis needs to be further improved. In this work, two kinds of serological biomarker species, microRNA and protein biomarker, have been detected simultaneously to identify HCC. Herein, a dual-aptamer hairpin DNA oligonucleotide is designed as the electrochemical sensing probe (ESP) to achieve this goal. The hairpin-structured DNA probe consists microRNA-16 (miR-16) complementary sequence and alpha fetoprotein (AFP) aptamer sequence, so it can both capture miR-16 and AFP. Once it hybridizes with miR-16, the hairpin structure is unlocked so that the terminal modified signal molecule (methylene blue, MB) would give a decreased electrochemical signal. Meanwhile, once it recognizes AFP, concanavalin A (ConA) modified silver nanoparticles (AgNPs) can bind to AFP at the sensing surface. An obvious electrochemical signal of AgNPs can thus be generated for AFP detection. In this way, one-step and simultaneous detection of miRNA-16 and AFP can easily be realized by collecting the two sensitive and non-interfering electrochemical signals. Compared with traditional single biomarker detection methods, this assay strategy can improve the accuracy of HCC by monitoring two kinds of serological biomarkers species. Besides, this novel electrochemical biosensor based on ESP is simple, low-cost and efficient, which make it promising to improve the accuracy and specificity for the diagnosis HCC in the future.

Keywords: Alpha fetoprotein (AFP); Electrochemical biosensor; Hairpin-structured DNA probe; Hepatocellular carcinoma biomarkers; microRNA-16.