摘要
目的：以重组麻风杆菌α２融合蛋白为抗原，建立酶联免疫吸附试验，即 α２－ ＥＬＩＳＡ。方法：用麻风病人和健康正常人的血清评价了α２－ＥＬＩＳＡ中有关试剂及最适浓度，其敏感性与ＰＧＬ－１－ＥＬＩＳＡ进行了比较。结果：（１）挥发性缓冲液醋酸铵碳酸盐（０．０１ ｍｏｌ／Ｌ）为α２抗原最佳包被液；（２）重组α２融合蛋白的最适包被浓度为５ μｇ／ｍｌ；（３）显色剂 ＯＰＤ的敏感性大于ＴＭＢ；（４）麻风病人血清中的抗体滴度分别为ｌｇＧ＞ｌｇＭ＞ｌｇＡ；结论：确定了α２－ＥＬＩＳＡ方法的最适实验条件，且该方法的敏感性与ＰＧＬ－１－ＥＬＩＳＡ相比无统计学差异，可望成为一种新的麻风血清学诊断和流行病学研究的方法。
To deve1op an enzyme-linked immunosorbent assay using a new recombinant M. leprae, a antigen(α2 - ELISA) for detecting the relevant antibodies in leprosy patients. Methods: In sera from leprosy patients (96)and normal Persons(50), the relevant reagents and their most optional condition of α2 - ELISA were evaluated, and the sensitivity of the new method was also compared with that of PGL- 1 ELISA. Results: (1)As a coating buffer to be used for α2- ELISA, Nh_4AC: CO_3 Would be most optional; (2) For M. leprae α2 antigen, 5 μg/ml was the most appropriate coating concentration; (3 ) In comparison with TMB, OPD was a more sensitive peroxidase substrate and (4) The titers of antibodies to α2 antigen in leprosy patients were IgG > IgM > IgA on an other basis. Conclusion: The serological activities of α2- ELISA are comparable with that of PGL- 1 ELISA and this method could be a new potential immunological tool in serodiagnosis and seroepidemiological investigation and, in particular, in identifying high risk group of leprosy infection.
出处
《中国麻风皮肤病杂志》
北大核心
1999年第2期12-14,共3页
China Journal of Leprosy and Skin Diseases
基金
国家教委科研基金
