抗-B淋巴细胞刺激因子自身抗体酶联免疫吸附法的建立和临床初步应用  

Development of enzyme-linked immunosorbent assay method for anti-B lymphocyte stimulator autoantibodies and its preliminary clinical application

作　　者：虞伟[1] 张燕[2] 李晓军[1] 武建国[1] 

机构地区：[1]南京军区南京总医院解放军临床检验医学研究所,江苏南京210002 [2]南京市鼓楼医院检验科,江苏南京210008

出　　处：《医学研究生学报》2007年第10期1014-1017,共4页Journal of Medical Postgraduates

基　　金：江苏省135工程医学重点学科基金资助项目(批准号:RC2002024);中国博士后科研基金资助项目(批准号:20060400940);江苏省博士后科研基金资助项目(批准号:0602008C)

摘　　要：目的:研究B淋巴细胞刺激因子抗体(BlyS-Ab)在临床相关疾病患者中的检出水平,探讨其在自身免疫性疾病中的作用。方法:优化实验反应条件,建立检测BlyS-Ab的酶联免疫吸附法(ELISA),并进行方法学考核和临床应用检测,研究血清BlyS-Ab水平与抗核抗体及其他自身抗体的关系。结果:BlyS-Ab精密度平均变异系数(CV值)为7.8%,特异阻断抑制试验最高抑制率达77.3%。临床研究结果显示,在系统性红斑狼疮(SLE)、类风湿性关节炎(RA)和重症肌无力患者中均有显著增高的BlyS-Ab阳性检出率,而在过敏性紫癜及慢性肾炎和肾病患者中亦发现有较高水平的BlyS-Ab存在,血清BlyS-Ab水平与抗核抗体(ANA)滴度呈高度正相关关系。在所测63例BlyS-Ab阳性患者中,ANA同时阳性者有59例,而64例BlyS-Ab阴性患者中,ANA阴性者为30例,两者检测结果具有显著的关联和中等程度的一致性(χ2=26.387,Kappa=0.404,P<0.001)。经与其他单特异性自身抗体对比研究发现,BlyS-Ab与抗双股脱氧核糖核酸(dsDNA)、抗干燥综合征A(SSA)、抗干燥综合征B(SSB)及抗核糖核蛋白(RNP)抗体亦存在一定程度的相关性(P<0.01或P<0.05)。结论:成功建立了人血清BlyS-Ab的ELISA测定方法,实验结果敏感、特异、可靠。血清BlyS-Ab检测可能有助于自身免疫性疾病的诊断。Objective：To investigate the prevalence of the antibodies against the B lymphocyte stimulator（BlyS）, （BlyS-Ab） in patients with clinical diseases, and to explore the role of BlyS-Ab in the pathogenesis of autoimmune diseases （AID）. Methods：By optimizing experimental conditions, we developed ELISA methods for serum levels of BlyS-Ab, followed by methodological and clinical application assessment and investigation of the relationship of serum BlyS-Ab levels with the titers of anti-nucleus antibodies （ANA） and other autoantibodies. Results：The intra-assay precision, namely the average coefficient values of BlyS-Ab was 7.8% , and the specific inhibition rate for BlyS-Ab by series concentration of BlyS was up to 77.3%. Clinical investigation revealed that the serum levels of BlyS-Ab was significantly higher in patients with autoimmune diseases （including SLE, RA, myasthenia gravis） and other medical cases（ such as allergic purpurics, chronic nephritis, etc. ） than in the normal control （P 〈0.01 ）. Significant positive correlation was found between the serum levels of BlyS-Ab and ANA titers. In 63 patients with BlyS-Ab positive, there were 59 with ANA positive, whereas in another 64 with BlyS-Ab negative, there were 30 with ANA negative, 60th with significant association and moderate coincidence （X^2 =26. 387, Kappa =0. 404,P 〈 0. 001 ）. To some extent, BlyS-Ab was also significantly associated with anti-dsDNA, anti-SSA, anti-SSB and anti-RNP antibodies （ P 〈 0.01 or P 〈 0.05） Conclusion：The ELISA method for human serum BlyS-Ab was successfully established, and the result of this assay was specific, sensitive and reliable. The detection of the serum levels of BlyS-Ab is potentially helpful to the diagnosis of autoimmune diseases.

关 键 词：B淋巴细胞刺激因子 自身免疫疾病 自身抗体 

分 类 号：R446.61[医药卫生—诊断学]