van der Sluijs Veer G, Moens H J
Eur J Clin Chem Clin Biochem. 1996 Nov;34(11):915-20. doi: 10.1515/cclm.1996.34.11.915.
A time-resolved immunofluorometric assay (TRIFMA) of autoantibodies to double-stranded DNA (dsDNA) is described. Biotinylated DNA was bound to the polystyrene solid phase coated with streptavidin. F(ab1')2 fragments from antibodies raised in rabbits to human IgG were labelled with Eu3+, and used in the assay to label the bound autoantibodies to dsDNA. The measuring range covers three decades. The proposed assay has good analytical properties. For calibration the First International Standard for antibodies to double-stranded DNA Wo/80 was used. The 97.5th percentile in normal persons is 20 kIU/l. Comparison of the TRIFMA and the Farr-assay in the analysis of 56 sera from 20 patients with systemic lupus erythematosus or lupus-like disease, 13 with other autoimmune diseases, and 10 blood-bank donors indicates a high degree of concordance (Kendall's tau = 0.56, p < 0.001). Further evaluation in 102 patients with systemic lupus erythematosus, lupus-like disease and other autoimmune diseases shows that sensitivity for active classical systemic lupus erythematosus is adequate, and specificity is excellent.
描述了一种用于检测抗双链DNA（dsDNA）自身抗体的时间分辨免疫荧光分析（TRIFMA）方法。生物素化的DNA与包被有链霉亲和素的聚苯乙烯固相相结合。用铕离子（Eu3+）标记兔抗人IgG抗体产生的F（ab1′）2片段，并用于该分析以标记与dsDNA结合的自身抗体。测量范围涵盖三个数量级。所提出的分析方法具有良好的分析性能。校准使用了抗双链DNA的第一国际标准品Wo/80。正常人的第97.5百分位数为20 kIU/l。对20例系统性红斑狼疮或狼疮样疾病患者、13例其他自身免疫性疾病患者以及10名血库供者的56份血清进行分析时，将TRIFMA与Farr分析进行比较，结果显示两者具有高度一致性（肯德尔tau系数=0.56，p<0.001）。对102例系统性红斑狼疮、狼疮样疾病和其他自身免疫性疾病患者的进一步评估表明，该方法对活动性典型系统性红斑狼疮的敏感性足够，特异性极佳。
