Zhang Shusheng, Du Ping, Li Feng
College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao 266042, China.
Talanta. 2007 Jun 15;72(4):1487-93. doi: 10.1016/j.talanta.2007.01.063. Epub 2007 Feb 9.
A new voltammetric enzyme-linked immunoassay system of 3,4-diaminobenzoic acid (DBA)-H(2)O(2)-horseradish peroxidase (HRP) for sensitive detection of prostate specific antigen (PSA) in human serum was present. In the proposed procedure, labelled HRP efficiently catalyzed the oxidation reaction of DBA by H(2)O(2) and generated the electroactive product, 2,2'-biamino-4,4'-bicarboxyl azobiphenyl, which produced a sensitive second-order derivative linear sweep voltammetric peak at potential of -0.62V (versus SCE) in Britton-Robinson (BR) buffer solution. The linear range for detection free HRP was from 4.0x10(-12) to 4.0x10(-9)gml(-1) and the detection limit was about 6.0x10(-13)gml(-1). The proposed new system could be used for detection of PSA ranging from 0.20 to 16.0ngml(-1) with a detection limit of 0.10ngml(-1), which was five times lower than that of the traditional o-phenylendiamine spectrophotometric enzyme-linked immunosorbent assay (ELISA) method. The proposed electrochemical ELISA method is simple, inexpensive, reproducible and sensitive, which shows potential for detecting PSA in clinical diagnosis.
提出了一种用于灵敏检测人血清中前列腺特异性抗原(PSA)的3,4-二氨基苯甲酸(DBA)-H₂O₂-辣根过氧化物酶(HRP)伏安酶联免疫分析新体系。在所提出的方法中,标记的HRP有效催化H₂O₂氧化DBA的反应,生成电活性产物2,2'-二氨基-4,4'-二羧基偶氮联苯,该产物在Britton-Robinson(BR)缓冲溶液中于-0.62V(相对于饱和甘汞电极)电位处产生灵敏的二阶导数线性扫描伏安峰。游离HRP的检测线性范围为4.0×10⁻¹²至4.0×10⁻⁹gml⁻¹,检测限约为6.0×10⁻¹³gml⁻¹。所提出的新体系可用于检测范围为0.20至16.0ngml⁻¹的PSA,检测限为0.10ngml⁻¹,比传统的邻苯二胺分光光度酶联免疫吸附测定(ELISA)方法低五倍。所提出的电化学ELISA方法简单、廉价、可重现且灵敏,在临床诊断中检测PSA方面显示出潜力。