基于巯基自组装膜的乙肝电化学免疫传感器

研制了以乙肝表面抗原(HBsAg)为检测对象的电化学免疫传感器.在金电极表面自组装半胱胺,戊二醛进行醛基化,最后通过竞争组装在自组装膜上同时固定乙肝表面抗体和硫堇,在PBS底液中以硫堇和辣根过氧化物酶(HRP)的还原峰电流的下降△/p(%)]对乙肝表面抗原进行定性和定量的检测.结果表明,传感器具有良好的选择性,丙肝阳性对照血清对其无干扰△/p随HBsAg的质量浓度(P)的增加而增大.HBsAg的质量浓度在0.85×10~(-2)～5.12×10~(-2)μg/mL时,线性方程为△/p=5.19+1.56×ρ,相关系数为0.981 5;HBsAg的质量浓度在0.32～1.28μg/mL时,线性方程为△/p=17.05+0.075×ρ,相关系数为0.990 6.并取临床血清进行检测,探索该传感器应用于临床检验的可行性.

The electrochemical immunosensor of hepatitis B based on self-assembled monolayers technology

The immunosensor was prepared by co-immobilizing thionine and Hepatitis B surface anti-body (HBsAb) on the gold electrode through covalently binding them to Au with a cysteamine/glutaral-dehyde linkage. Hepatitis B surface antigen (HBsAg) was detected qualitatively and quantificationally by the peak current decrease percentage (△/p%) of the thionine and horseradish peroxidase (HRP) in the PBS solution. The cyclic voltammngrams of the immunosenor displayed a couple of redox peaks current of the thionine. The peak current decreased after the HBsAg reacted with the HBsAb. Further more, the se-lectivity of the sensor was also studied, there's no peak current decrease while detecting the Hepatitis C antigen. The △/p% was linening relationship with the quality concentration of the HBsAg ranging from0.85×10~(-2)～5.12×10~(-2.)μg/mL and 0. 32 ～ 1.28μg/mL with the correlation coefficient of 0. 9815 and 0. 9906, respectively. The immunosensor was used in the clinical laboratory of human serum, which was in good agreement with the ELISA.