CAJ | 학술논문

目的:探索一种新的免疫传感器,以提高检测的敏感性及抗体滴度的线性范围.方法:将碳油墨和银、氯化银油墨滴在PET板,作为两电极测试条,碳为工作电极,用不同的方法把可溶性抗原固定在工作电极的末端,银、氯化银电极作为对照.在电化学工作站中,采用循环伏安法和示差脉冲伏安法测试抗体的效价,并对这些方法的效果进行比较.在此基础上开发2种固定日本血吸虫抗原的印刷电极免疫传感器:戊二醛交联(GA)传感器和壳聚糖戊二醛(Chit-GA)传感器,并采用循环伏安法和示差脉冲伏安法测试抗体滴度检测范围.结果:GA膜修饰电极的日本血吸虫抗原最佳包被浓度为50 μg/L,Chit-GA膜修饰电极的最佳浓度值为10 μg/L；2种电极的免疫反应时间均为1 min; GA,Chit-GA免疫传感器检测范围分别为1∶1000～1∶400,1∶1000～1∶ 500.示差脉冲伏安法电流的测试值会随着血清效价增大而增大.结论:GA传感器和Chit-GA的交联传感器具有较高的检测灵敏度,反应时间短,检测抗体滴度线性范围宽,并且与日本血吸虫抗体滴度具有良好的线性关系.
목적:탐색일충신적면역전감기,이제고검측적민감성급항체적도적선성범위.방법:장탄유묵화은、록화은유묵적재PET판,작위량전겁측시조,탄위공작전겁,용불동적방법파가용성항원고정재공작전겁적말단,은、록화은전겁작위대조.재전화학공작참중,채용순배복안법화시차맥충복안법측시항체적효개,병대저사방법적효과진행비교.재차기출상개발2충고정일본혈흡충항원적인쇄전겁면역전감기:무이철교련(GA)전감기화각취당무이철(Chit-GA)전감기,병채용순배복안법화시차맥충복안법측시항체적도검측범위.결과:GA막수식전겁적일본혈흡충항원최가포피농도위50 μg/L,Chit-GA막수식전겁적최가농도치위10 μg/L；2충전겁적면역반응시간균위1 min; GA,Chit-GA면역전감기검측범위분별위1∶1000～1∶400,1∶1000～1∶ 500.시차맥충복안법전류적측시치회수착혈청효개증대이증대.결론:GA전감기화Chit-GA적교련전감기구유교고적검측령민도,반응시간단,검측항체적도선성범위관,병차여일본혈흡충항체적도구유량호적선성관계.
Objective:To improve the sensitivity and the linear range of electrochemical immunosensor to detect Schistosoma japonicum (S.japonicum) antibody.Methods:Carbon inks and silver/silver chloride inks were printed on a polyethylene terephthalate (PET) board to make a two-electrode test strip,where carbon was the working electrode and S.japonicum soluble egg antigen (SEA) was fixed at one end of working electrode by different methods; silver/silver chloride electrode was used as control.We tested the valency of the antibody by cyclic voltammetry (CV) and differential pulse voltammetry (DPV) in an electrochemistry workstation,and conducted comparison with the results of ELISA.Two new immunosensing electrodes have been developed,based on glutaraldehyde cross-linked (GA) or chitosan-glutaraldehyde cross-linked (Chit-GA) transducer fixing S.japonicum antigen.We tested the titer of the antibody by means of CV and DPV.Results:Our experimental S.japonicum antigen (50 μg/L) is the optimal test concentration for the GA sensor,and 10 μg/L for Chit-GA sensors.The immune reaction time of both electrodes is all essentially complete in 1 minute.The linear range for S.japonicura antibody in human positive serum sample detection by the glutaraldehyde cross-linked immunosensor is 1∶1000 to 1∶400,and by the chitosan-glutaraldehyde cross-linked immunosensor is 1∶1000 to 1∶500.As the concentration of dilution ratio of S.japonicum antibody in human positive serum sample increased,the test value of DPV increased proportionally.Conclusion:GA sensor and Chit-GA cross-linked S.japonicum sensors have high sensitivity and broad linear range response,and both exhibited a good linear relationship between the DPV signal and the test antibody titer.