磁分离技术和纳米金比色法用于嗜碱性粒细胞活化试验研究

Basophil activationtest (BAT) is a safe and efficient technique for the diagnosis of allergic diseases, which is one of the main methods for the clinical diagnosis of allergic diseases in vitro. Flow cytometry has been mainly used in BAT. However, this method showed some disadvantages, such as long detection time, cumbersome operation and high cost. To solve these problems, we developed a magnetic bead separation system with surface modified HLA-DR antibody. After co-incubation with peripheral blood of allergic patients for 5 min, cells with HLA- DR markers on the surface were seprated by magnetic separation technology. The results showed that the proportion of basophil cells in whole blood increased from 0.2% to 11.5% after screening, which effectively improved the sensitivity of BAT. Further, we developed a gold nanoparticle- based colorimetry with the CD63 aptamer modification on the surface of gold nanoparticles. Due to the specificity identification of CD63 aptamer with CD63 antibody after activation of basophils, the gold nanoparticles can gather together on the surface of basophils, thus resulting in the color changing from red to violet, which finally realized visual colorimetric detection of the activation of basophils. We herein believe our strategy will provide a new method for clinical BAT.