ELISA法检测可溶性人类白细胞抗原-Ⅰ及广东人参考值的测定

Detection of soluble HLA classⅠantigens by ELISA and determination of its reference value in population of Guangdong province

目的：探讨可溶性人类白细胞抗原-I(sHLA-I)的定量检测技术，并用于检测广东地区人群的参考值。方法：以W6/32包被酶标板，捕捉样品中sHLA-I，β2微球蛋白抗体为一抗，再加酶标二抗及底物显色。根据不同浓度标准sHLA-I显色后Dλ值的标准曲线，检测60例正常广东人的sHLA-I含量。结果：sHLA-I最低检测限为2.84ng/ml，批内变异系数为5.80％，批间变异系数为9.00％，回收率为98.5％-100.15％。广东人sHLA-I正常值为（699.54±360.10）ng/ml。结论：ELISA法检测sHLA-I具有灵敏度高、特异性强、稳定性好等优点。

Objective To explore a method for quantitative detection of soluble human leukocyte antigen class Ⅰ(HLA-Ⅰ) antigens, with the assistance of which we attempt to define the reference value of the antigen in the population of Guangdong Province. Methods Sandwich enzyme-linked immunosorbent assay (ELISA) was employed with W6/32 monoclonal antibody serving as the solid phase antibody and β2 microglobulin antibody as the first antibody. HRP-labeled second antibody and the substrate were added for enzyme-catalyzed coloring. On the basis of the standard curve obtained by sHLA-Ⅰstandard reagent in serial dilution, the concentration of sHLA-I antigens in the samples of 60 normal subjects from the population of Guangdong province was detected. Results The sensitivity of this assay was 2.84 ng/ml with variation coefficients of 5.80% within the same batch and 9.00% between batches. The recovery rate ranged from 98.57% to 100.15%. The level of sHLA-Ⅰantigens in normal individuals was 699.54±360.10 ng/ml. Conclusion Sandwich ELISA is sensitive, specific and stable in the detection of sHLA-Ⅰ.