关键词: 流感嗜血杆菌, P6蛋白, 单克隆抗体, 胶体金免疫层析法

Abstract: To establish a rapid and sensitive detection method for Haemophilus influenzae, mice were immunized with P6 recombinant protein, and hybridoma cells were prepared by monoclonal antibody technology. Monoclonal antibodies were obtained by ascites preparation and affinity purification. An immunochromatographic test paper was developed based on the double antibody sandwich principle. As a result， two monoclonal antibodies against P6 recombinant protein were successfully screened and prepared, and their titers reached 1∶ 512 000. It confirmed that the monoclonal antibodies could specifically recognize the natural P6 protein of Haemophilus influenzae by Western Blot. The KD values of the two monoclonal antibodies were 10-9 and 10-10, respectively, and the affinity was good. By the established approach， Haemophilus influenzae was quickly and specifically detected in 10 min, and it had no cross-reaction with 9 common respiratory pathogens. After accelerated stability test, it could be stored and used at room temperature for one year, with good repeatability and stability.

Key words: Haemophilus influenzae, P6 protein, monoclonal antibody, colloidal gold immunochromatograph