桥式生物素-亲合素标记提高藻胆蛋白免疫荧光法灵敏度的研究

Study on bridged avidin biotin technique improving the sensitivity of phycobiliprotein immunofluorescence assay

目的提高藻胆蛋白免疫荧光法检测的灵敏度。方法在藻胆蛋白免疫荧光法(PBPIFA)的检测流程中，引入桥式亲合素－生物素系统(BRAB)，称为BRAB-PBPIFA,用ELISA法与藻胆蛋白免疫荧光直接法和桥式BAS法，对定值的标准质控抗原HBsAg样品及500份血清样品同时进行测定。结果BRAB应用于藻胆蛋白免疫荧光法中，能显著提高该法对低抗原含量样品的检出，对弱阳性血清的检出率可提高1.5倍。无蛋白封闭试剂Tween 20在BRAB-PBPIFA中具有较好的封闭效果。结论BRAB标记可提高藻胆蛋白免疫荧光法检测的灵敏度。

Aim To improve the sensitivity of phycobiliprotein immunofluorescence assay(PBPIFA), by introducing into bridged avidin biotin(BRAB) technique,known as BRAB-PBPIFA. Methods Using ELISA as a gold standard, effectiveness of three assays(direct PBPIFA, BRAB-PBPIFA and ELISA)were compared in simultaneous detection of standard antigen and 500 serum samples. Results BRAB improved greatly sensitivity of PBPIFA in detecting antigen of low concentration, and detection rate of weakly positive samples could increase by one times and a half. In the applying, protein-free blocking agent Tween 20 to BRAB-PBPIFA,possessed better blocking effect. Conclusion Combination of BRAB technique with PBPIFA can improve the detcctive sensitivity.