一种梅毒螺旋体重组抗原胶体金的制备

目的制备一种可以特异、准确的检测血清中梅毒螺旋体抗体的胶体金免疫层析试纸。方法利用原核表达系统构建一种包含梅毒螺旋体多个免疫表位的重组抗原B1,采用ELISA的方法对重组抗原进行活性检测。胶体金标记重组抗原B1,制备胶体金免疫层析试纸。结果重组抗原B1在BL21表达载体中有较高表达,ELISA结果显示B1与梅毒患者血清能特异、敏感的结合,与乙型肝炎病毒血清及阴性血清无交叉反应。应用B1蛋白制备的胶体金诊断试纸检测100份梅毒标准血清,结果均为阳性,与乙型肝炎患者血清及健康人群血清无交叉反应。结论梅毒螺旋体胶体金快速诊断试纸具有良好特异性和敏感性,可以快速、准确的诊断梅毒螺旋体的感染。

Development of the Dipstick of Gold-Immuno Chromayography Assay for the Diagnosis of Treponema

Objective To develop the dipstick of gold immunochromatograpy assay(GICA) with specific and sensitive for the detection the antibody of treponema. Methods The recombinant B1 with multi-epitopes derived from treponema was constructed by prokaryotic expression system. Indirect ELISA was carried out to test the immunocompetence of B1. The colloidal gold labeled protein B1 was assembled to the GICA dipstick. Results A high level expression of the recombinant protein could be seen in the BL21 expression vector,the result of ELISA indicates that the protein B1 could be binded with the standard serum of treponema,but no cross-reaction with the serum samples of hepatitis B virus and negative serum. The testing results showed that ICA method with 100 % sensitivity in 100 positive samples. However,no cross reaction with other serum could be seen. Conclusion The method of treponema ICA is specificity and sensitivity. It can be used for the diagnosis of treponema quickly and accurately.