Blount S, Griffiths H, Emery P, Lunec J
Department of Rheumatology, University of Birmingham, England, UK.
Clin Exp Immunol. 1990 Sep;81(3):384-9. doi: 10.1111/j.1365-2249.1990.tb05343.x.
During the development of an ELISA to measure anti-DNA antibodies in systemic lupus erythematosus (SLE) sera, native dsDNA was found not to be the most appropriate antigen to use in ELISA assays for differentiating between SLE patients and those with rheumatoid arthritis (RA), a disease also associated with circulating serum anti-DNA antibodies. By modifying the ELISA technique to incorporate human DNA, denatured by reactive oxygen species, to detect anti-DNA antibodies in SLE sera, results consistently showed an increase in antibody binding when compared with the native antigen; no such trend was observed in the comparable group of RA patients. Using this assay serum anti-dsDNA antibody levels were measured in a group of 20 controls, 20 RA patients (10 seropositive and 10 seronegative) and 30 SLE patients (15 with clinically active disease, 15 with inactive disease). A comparison with the standard radioimmunoassay used to measure anti-DNA antibodies for the diagnosis of SLE showed that the ELISA assay using modified DNA performed better than the standard radioimmunoassay offering an improvement in both clinical specificity and sensitivity. The improved method particularly reduced the problem of false-negative results for SLE patients shown clinically to be either mildly active or inactive.
在开发用于检测系统性红斑狼疮(SLE)血清中抗DNA抗体的酶联免疫吸附测定(ELISA)过程中,发现天然双链DNA(dsDNA)并非ELISA检测中区分SLE患者与类风湿性关节炎(RA)患者(一种也与循环血清抗DNA抗体相关的疾病)的最合适抗原。通过改进ELISA技术,加入经活性氧变性的人类DNA来检测SLE血清中的抗DNA抗体,结果始终显示与天然抗原相比抗体结合增加;在可比的RA患者组中未观察到这种趋势。使用该检测方法,对20名对照、20名RA患者(10名血清阳性和10名血清阴性)以及30名SLE患者(15名临床活动期患者和15名非活动期患者)的血清抗dsDNA抗体水平进行了测量。与用于诊断SLE的标准放射免疫测定法相比,使用改良DNA的ELISA检测表现更好,在临床特异性和敏感性方面均有提高。改进后的方法尤其减少了临床显示为轻度活动或非活动的SLE患者假阴性结果的问题。