Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan 81746-73441, Iran.
Department of Biotechnology, Faculty of Advanced Sciences and Technologies, University of Isfahan, Isfahan 81746-73441, Iran.
J Immunol Methods. 2018 Jul;458:26-32. doi: 10.1016/j.jim.2018.04.001. Epub 2018 Apr 12.
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 FeO and AlO 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-FeO) as the capture antibody (Ab). In addition, another hepatitis B antibody against different epitope of HBsAg (Ab) and glucoamylase both were immobilized on AlO nanoparticles. After formation of the sandwich immune complex between Ab and Ab immobilized on S-FeO and AlO 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 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.
乙型肝炎病毒(HBV)感染是全球主要的公共卫生问题,也是导致死亡的主要原因。针对这种感染,最重要的诊断标志物是乙型肝炎表面抗原(HBsAg)。在本研究中,设计并研究了一种新颖、廉价、便携且灵敏的基于功能化 FeO 和 AlO 纳米粒子的 ELISA 方法,用于诊断 HBsAg,其检测策略是使用便宜且易得的个人血糖仪(PGM)来检测葡萄糖浓度。该 ELISA 系统使用固定在链霉亲和素包被的磁性氧化铁颗粒(S-FeO)上的针对 HBsAg 的乙型肝炎抗体作为捕获抗体(Ab)。此外,另一种针对 HBsAg 不同表位的乙型肝炎抗体(Ab)和糖化酶都固定在 AlO 纳米粒子上。通过 HBsAg 分别在 S-FeO 和 AlO NPs 上固定的 Ab 和 Ab 形成三明治免疫复合物后,使用糖化酶将淀粉转化为葡萄糖。然后,使用 PGM 测量葡萄糖浓度。根据 HBsAg 浓度与葡萄糖浓度之间的线性关系计算 HBsAg 的浓度。在最佳条件下,该测定法对“ay”和“ad”亚型的 HBsAg 的检测限分别为 0.3 至 0.4 ng/ml。结果表明,该测定法在灵敏度、现场、特异性、成本、简单性、便携性和重现性方面可与商业试剂盒相媲美。该方法可用于世界上的贫困地区和输血中心。据我们所知,这是首次使用 PGM 检测 HBSAg 的报道。