使用重组抗原通过酶联免疫吸附测定法检测抗PL - 12自身抗体；免疫反应区域的研究
Detection of anti-PL-12 autoantibodies by ELISA using a recombinant antigen; study of the immunoreactive region.
作者信息
García-Lozano J R, González-Escribano M F, Rodríguez R, Rodriguez-Sanchez J L, Targoff I N, Wichmann I, Núñez-Roldán A
机构信息
Servicio de Inmunología, Hospital Universitario Virgen del Rocío, Sevilla, Spain.
出版信息
Clin Exp Immunol. 1998 Nov;114(2):161-5. doi: 10.1046/j.1365-2249.1998.00720.x.
Autoantibodies to aminoacyl-tRNA synthetases are highly associated with myositis and detection is important in clinical diagnosis; however, current methods of screening limit its clinical utility. In the present study, alanyl-tRNA synthetase (PL-12) recombinant protein was obtained by immunological screening of a HeLa expression library and used in an ELISA with 22 anti-PL-12 sera, 200 autoimmune sera negative for PL-12 and 100 healthy individual sera. Sensitivity of the method was 95% (21/22) and specificity 100%. Mapping of the immunoreactive region was carried out using three anti-PL-12 sera and different recombinant protein-derived peptides. Results show that the same conformational epitope located within amino acids 730-951 of the PL-12 antigen outside the catalytic region was recognized by the three anti-PL-12 sera tested. We conclude that ELISA using recombinant protein is an effective and useful method for routine screening for anti-PL-12 autoantibodies.
氨酰-tRNA合成酶自身抗体与肌炎高度相关，其检测在临床诊断中具有重要意义；然而，目前的筛查方法限制了其临床应用。在本研究中，通过对HeLa表达文库进行免疫筛选获得了丙氨酰-tRNA合成酶（PL-12）重组蛋白，并将其用于酶联免疫吸附测定（ELISA），检测对象包括22份抗PL-12血清、200份PL-12阴性的自身免疫血清以及100份健康个体血清。该方法的灵敏度为95%（21/22），特异性为100%。使用三份抗PL-12血清和不同的重组蛋白衍生肽对免疫反应区域进行了定位。结果显示，所检测的三份抗PL-12血清识别的是位于PL-12抗原催化区域外730-951氨基酸内的相同构象表位。我们得出结论，使用重组蛋白的ELISA是一种用于抗PL-12自身抗体常规筛查的有效且实用的方法。
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