Virus Model Of Multiple High-throughput Screening Of Blood Flow Fluorescence Microsphere Technique Establishment And Testing Of Blood Infected With Hbv, Hcv, Applied Research,

Nowadays, the phenomenon of infecting HBV, HCV, HIV and TP after blood transfusion is prevalent and payed much attention by the society. The most important task we are facing is to strictly control blood screening. Conventional ELISA still has its detecting limits, eg: low-throughput, time-wasting and low sensitivity. In this research, Luminex fluorescent microsphere technology is used, and combined with sandwich capture method, indirect capture method and biotin-avidin system, in order to establish a high-throughput, fast and sensitive method to detect HBsAg, HCV-Ab and other infection factors in blood.This study is divided into three parts:1. Optimizing biotinylation method.2. Optimizing coupling method of antigen/antibody to fluorescent microsphere.3. Using Luminex fluorescent microsphere technology to detect HBsAg andHCV-Ab in human serum.The results show that PBS and DMF are better than NaHCO₃ and DMSO for biotinylation of antibody. The biological activity and the biotinylation efficiency of antibody keeps well after biotinylation. It is better for antigen in 100mM(pH6.0)MES and antibody in 50mM(pH5.0)MES to couple with fluorescent microsphere, and the best consentration of antigen/antibody is 100 u g/ml.The sensitivity and specificity of microsphere are both 100% in detection HBsAg of 128 HBV serum, while 66.67% and 71.43% by ELISA. The sensitivity of microsphere is 89.19% in detection HCV-Ab of 74 HCV serum. The conclusion is that fluorescent microsphere technology is better than ELISA.