B细胞激活因子水平ELISA检测方法的建立及临床初评

An ELISA method for determination of serum B lymphocyte stimulator levels and its clinical assessment

目的:建立血清B细胞激活因子(B lymphocyte stimulator,BlyS)水平的ELISA检测方法,并初步探讨其临床价值。方法:以商品化的抗人BlyS单克隆抗体、生物素化抗人BlyS多克隆抗体,采用双抗夹心法自行建立BlyS血清水平的ELISA检测方法;并对80例风湿性疾病患者［50例系统性红斑狼疮(SLE)、20例类风湿关节炎(RA)、10例干燥综合征(SS)］、60例自身免疫性肝病患者［35例原发性胆汁性肝硬化(PBC)、25例自身免疫性肝炎(AIH)］和40例正常人的血清BlyS水平进行检测。结果:成功建立血清BlyS水平的ELISA检测法且重复性较好,80例风湿性疾病患者的血清BlyS水平为(8.23±1.42)ng／ml,60例自身免疫性肝病患者为(3.40±0.79)ng／ml,40例正常人的血清BlyS水平为(3.19±0.88)ng／ml。与正常人相比,风湿性疾病组的BlyS血清水平明显升高(P<0.01),而自身免疫性肝病组的BlyS血清水平升高则不明显。结论:本研究所建立的BlyS血清水平ELISA检测方法简便、可靠、准确,为后续研究BlyS与自身免疫性疾病尤其是风湿性疾病的关系奠定了基础。

Objective: To establish an ELISA method for determining the serum levels of B lymphocyte stimulator (BlyS) and to evaluate its clinical significance. Methods: Anti-human BlyS monoclonal antibody and biotinylated anti-human BlyS polyclonal antibody were used to detect serum BlyS levels by ELISA. Serum samples from 80 systemic immune-based rheumatic disease patients (50 with systemic lupus erythematosus [SLE],20 with rheumatoid arthritis [RA],10 with primary Sjogren's syndrome [SS]), 60 with autoimmune liver disease (ALD) patients (35 with primary biliary cirrhosis [PBC],25 with autoimmune hepatitis [AiH]) were assayed for BlyS levels, and the results were compared with those of 40 healthy controls. Results: The median level of serum BlyS was (8. 23±1. 42) ng/ml in rheumatic disease patients, (3. 40±0. 79) ng/ml in ALD patients, and (3. 19±0. 88) ng/ml in healthy controls. The serum levels of BlyS in rheumatic disease patients were significantly higher than those in the normal cohort (P<0. 01), while the levels in ALD patients showed no significant differences. Conclusion: ELISA assay established in this study is reliable, simple and has precise results, which might be used for later study on BlyS and autoimmune diseases, such as rheumatic diseases.