山西汉族可溶性HLA-Ⅰ类抗原检测

Detection of serum-soluble HLA class Ⅰ antigens in the population of the Han nationality in Shanxi province

目的:定量检测正常人血清中可溶性人白细胞抗原-Ⅰ(sHLA-Ⅰ),以探讨山西汉族人群的正常参考值。方法:ELISA法检测血清sHLA-Ⅰ类抗原水平。将不同浓度的标准品和待检血清分别加入包被有特异性sHLA-Ⅰ抗体的酶标板,与生物素化的sHLA-Ⅰ、辣根过氧化物酶(HRP)标记的链霉亲和素共同孵育,弃上清并用洗涤液洗涤,加底物应用液四甲基联苯胺(TMB)显色,以酶标仪450nm测定吸光度值,根据不同浓度标准品显色后测得的吸光度值绘制标准曲线,测定60例健康山西人的sHLA-Ⅰ含量。结果:ELISA法能够准确检测sHLA-Ⅰ含量,60例健康山西人的sHLA-Ⅰ含量为(382.62±106.68)ng.ml-1。结论:山西汉族人群sHLA-Ⅰ正常值为(382.62±106.68)ng.ml-1。

Objective： To evaluate the reference value of soluble human leukocyte antigen class I （sHLA- I ） in the population of the Han nationality in Shanxi province. Methods： Enzyme-linked immunosorbent assay（ELISA） was used to measure the level of sHLA- I. Microtitration wells were coated with sHLA- I antibody of defined and unique epitope specificity. Standards and corre- sponding samples were added into different mierotitration wells respectively, and then biotinylated sHLA- I and streptavidin labeled with enzyme horseradish peroxidase（HRP） were added into each well. After incubation and washing, a TMB substrate Solution was added to the wells and color developed in proportion to the amount of sHLA- I bound. The intensity of the color was measured at 450nm. Standard curve was obtained by quantification of sHLA- I standard reagent in series dilution, the level of sHLA- I antigens was examined in 60 normal persons. Results： sHLA- I antigens can be exactly quantified by ELISA, and the average concentration ob- tained from 60 normal persons was （382.62,106. 68）ng· ml-1. Condnsions：The level of sHLA- I antigens in normal individuals of the Han nationality in Shanxi province was （382. 62±106. 68） ng· ml-1.