Herein we designed a novel, highly sensitive, simple and amplified fluorescence immunosensing strategy for hepatitis B virus surface antigen (HBV surface antigen) (HBsAg) as a model based on the construction of a sandwich type probe. The operation mechanism of this immunosensing strategy is implemented by capturing and then stimulation-based-releasing of entrapped dye in the fluorescent capsules. The proposed probe is made by the Fe3O4 magnetic nanoparticle (Fe3O4 MNP) as a probe collector site and the Rhodamine B loaded-mesoporous silica nanoparticle (MSN-Rh.B) as a fluorescent mesoporous capsule and signal amplifier site. Such a methodology is benefited, from the advantages of the high ability of MSNs to be used as a scaffold for efficient dye encapsulation and the magnetic nanoparticles as efficient biological carriers. Under optimal conditions, the fluorescence signal (The fluorescence of solutions was measured using a quartz fluorescence cell (PMT voltage:720, Ex wavelegth:540, Em wavelength:568, All measurements were carried out at room temperature) increased with the increment of HBsAg concentration in the linear dynamic range of 6.1 ag/ml to 0.012 ng/ml with a detection limit (LOD) of 5.7 ag/ml. The relative standard deviation, measured between the resulting fluorescence peaks was obtained by 6.0%
