Highly sensitive solution immersed silicon (SIS) biosensors were developed for detection of hepatitis B virus (HBV) infection in the early stage. The ultrasensitivity for overlayer thickness at the nonreflecting condition for the p-polarized wave is the basis of SIS sensing technology. The change in thickness due to biomolecular interactions and change in refractive index of the surrounding buffer medium were assessed simultaneously using two separate ellipsometric parameters (Ψ and Δ), respectively, from a single sensing spot. A direct antigen-antibody affinity assay was used to detect and quantify hepatitis B surface antigen (HBsAg), which is the early stage biomarker for HBV infection. The detection limit of 10 pg/ml was achieved for HBsAg in the human blood serum, which is comparable with the results of enzyme-linked immunosorbent assay and other hybrid assays. The SIS sensor's response time was less than 10 min. The SIS sensors exhibit excellent stability and high signal-to-noise ratio, and are cost-effective, which makes them a suitable candidate for point-of-care applications.