Houtman P M, Kallenberg C G, Limburg P C, Huitema M G, van Rijswijk M H, The T H
Clin Exp Immunol. 1985 Dec;62(3):696-704.
We describe a solid-phase enzyme-linked immunosorbent assay (ELISA) for quantitation of antibodies to nucleoribonucleoprotein (nRNP/Sm). nRNP/Sm was purified from rabbit thymus acetone powder by immunoaffinity chromatography and characterized by counterimmunoelectrophoresis (CIE) and immunoblotting using sera with well-known specificities. The purified antigen was used in ELISA. Positive results in ELISA were obtained only in sera with anti-nRNP or anti-Sm specificity as determined in CIE. Levels of anti-nRNP/Sm as quantitated by ELISA were higher in the sera of patients with active connective tissue disease (n = 7) than in those with inactive disease (n = 6) (P less than 0.01). Differences in anti-nRNP/Sm levels were also found between patients with mildly active disease (n = 19) and those with active disease (P less than 0.01). Fluctuations of anti-nRNP/Sm levels related to disease activity were seen in longitudinal observation. Although anti-nRNP/Sm levels as quantitated by ELISA correlated with titres of antinuclear antibodies as determined by immunofluorescence (r = 0.46, P less than 0.05), quantitation of anti-nRNP/Sm by ELISA is superior since the assay is antigen-specific and its quantitation independent of titration-related inaccuracies.
我们描述了一种用于定量检测抗核核糖核蛋白(nRNP/Sm)抗体的固相酶联免疫吸附测定(ELISA)方法。nRNP/Sm通过免疫亲和层析从兔胸腺丙酮粉中纯化得到,并通过对流免疫电泳(CIE)以及使用具有已知特异性的血清进行免疫印迹进行鉴定。纯化后的抗原用于ELISA检测。ELISA检测的阳性结果仅在CIE检测中具有抗nRNP或抗Sm特异性的血清中获得。ELISA定量检测的抗nRNP/Sm水平在活动性结缔组织病患者(n = 7)的血清中高于非活动性疾病患者(n = 6)(P < 0.01)。在轻度活动性疾病患者(n = 19)和活动性疾病患者之间也发现了抗nRNP/Sm水平的差异(P < 0.01)。在纵向观察中可见抗nRNP/Sm水平与疾病活动度相关的波动。虽然ELISA定量检测的抗nRNP/Sm水平与免疫荧光法测定的抗核抗体滴度相关(r = 0.46,P < 0.05),但ELISA法检测抗nRNP/Sm更具优势,因为该检测具有抗原特异性,且其定量不受滴定相关误差的影响。