We report the first demonstration of bovine serum albumin (BSA)-dual-probe sensing platform for B/C genotyping of hepatitis B virus based on the relationship between probe length and melting temperature. With the specially designed probes of genotypes B and C from HBV, this proposed BSA-dual-probe sensing platform is incubated in the same sample solution at two temperatures sequentially, accompanied by each electrochemical measurement. According to the relationship of two amperometric signals, the genotype of HBV can be successfully distinguished. The proposed genotyping sensor provides a novel strategy for HBV genotypes B and C with its advantage of simplicity, rapidness, reproducibility, and accuracy. It opens up a new avenue for genotyping based on the biomolecular characteristics of oligonucleotide. (C) 2016 Elsevier B.V. All rights reserved.