Ait Kaci A A, Monier J C
Pathol Biol (Paris). 1982 Feb;30(2):73-8.
The present paper describes a new ELISA type immunoenzymatic method for the detection of antibodies directed against nDNA, dDNA, and double stranded RNA (poly A, poly U and poly l, poly C). The antigens are adsorbed on glass beads but as some nucleic acids are difficult to adsorb on glass beads, polylysine was used to bind the antigens to the beads. The beads were saturated with sheep serum to get rid possible unspecific bindings. The sera are diluted before use with PBS containing sheep serum. The rate of antinucleic acid antibodies is obtained by subtraction of OD with antigen from OD without antigen. The technique was applied to human sera from patients with various autoimmune diseases (mostly disseminated lupus erythematosus). There is a rather good correlation between the determinations of anti-nDNA antibodies obtained with the ELISA and with indirect IF on Crithidia Luciliae.
本文描述了一种新型酶联免疫吸附测定(ELISA)免疫酶法,用于检测针对天然DNA(nDNA)、变性DNA(dDNA)和双链RNA(聚腺苷酸、聚尿苷酸以及聚肌苷酸、聚胞苷酸)的抗体。抗原吸附在玻璃珠上,但由于一些核酸难以吸附在玻璃珠上,因此使用聚赖氨酸将抗原与珠子结合。珠子用羊血清饱和以消除可能的非特异性结合。血清在使用前用含羊血清的磷酸盐缓冲盐水(PBS)稀释。抗核酸抗体的比率通过从不含抗原的光密度(OD)中减去含抗原的OD来获得。该技术应用于患有各种自身免疫性疾病(主要是播散性红斑狼疮)患者的人血清。用ELISA法和利什曼原虫间接免疫荧光法检测抗nDNA抗体的结果之间有相当好的相关性。