Background: To develop a equipment free, and can be widely used in clinical practice biosensor-based microarray for hepatitis B virus Pre-C/BCP mutation assay. Methods: A thin film optical biosensor were applied for amplification the microarray signal in situ. And HBV sites 1762, 1764,1768,1770 and 1896 were selected as the targets and the microarray were be fabricated. The 5 mutated plasmids contained 1762, 1764,1768,1770 and 1896 sites and 30 HBV sera were be tested in our study and all the plasmids and sera PCR products were be assayed by really time PCR and sequencing. Results: