A novel method for sensitive, low-cost and portable detection of hepatitis B surface antigen using a personal glucose meter

Hepatitis B virus (HBV) infection is the major public health problem leading cause of death worldwide. The most important diagnostic marker for this infection is hepatitis B surface antigen (HBsAg). In this study, a novel, inexpensive, portable and sensitive ELISA method was designed and investigated for diagnosis of HBsAg based on the functionalized Fe₃O₄ and Al₂O₃ nanoparticles, with the strategy for detecting the concentration of glucose using a cheap and accessible personal glucose meter (PGM). The ELISA system was constructed using hepatitis B antibody against HBsAg immobilized on streptavidin coated magnetic iron oxide particles (S-Fe₃O₄) as the capture antibody (Ab₁). In addition, another hepatitis B antibody against different epitope of HBsAg (Ab₂) and glucoamylase both were immobilized on Al₂O₃ nanoparticles. After formation of the sandwich immune complex between Ab₁ and Ab₂ immobilized on S-Fe₃O₄ and Al₂O₃ NPs, respectively, through HBsAg, starch was converted into glucose using glucoamylase. Then, the glucose concentration was measured using PGM. The concentration of HBsAg was calculated based on the linear relation between the concentrations of HBsAg and glucose. Under optimal conditions, this assay showed detection limit values of 0.3 to 0.4 ng ml⁻¹ for “ay” and “ad” subtypes of HBsAg, respectively. The results indicate that the designed assay is comparable to the commercial kits in terms of sensitivity, on-site, specificity, cost, simplicity, portability and reproducibility. The presented method can be used in disadvantaged areas of the world and blood transfusion centers. To the best of our knowledge, this is the first report of using PGMs for HBSAg detection.