摘要
用牛血清AGE(BSA AGE)作为包被抗原 ,采用对氧化糖基化产物———Nε 羧甲基赖氨酸 (CML)特异的抗体 (抗KLH AGE)作为检测抗体 ,建立竞争ELISA检测系统。将该法和荧光分光光度法所测AGE水平进行比较 ,并对 117例长期血液透析 (HD)病人的血清AGE水平进行定量观察。结果显示 :ELISA法检测血清AGE的最低检测限为 0 .1μg/ml,平均批内变异系数 (CV)为 2 .2 % ,批间CV为 9.0 % ,回收率为 97%～ 10 7%。用该法测定的 117例血透病人血清AGE水平中位数为 0 .5 μg/ml,而年龄匹配的正常人中仅有 9.4%的血清AGE含量>0 .1μg/ml。同时用ELISA法和荧光分光光度法检测 44例病人血清AGE ,两种方法所得结果相关非常显著 (P<0 .0 1)。上述结果表明 ,竞争ELISA法测定人血清AGE水平 ,具有方法简便、准确、特异性高和成本低的优点 ,符合临床使用要求 ,可作为反映病人氧化糖基化损伤的一种检测方法。
A solid phase, enzyme linked immunosorbent assay was developed to measure human serum AGE, and to elucidate its application in hemodialysis(HD) patients. Competitive ELISA using BSA AGE as the coating antigen and the specific antibody against CML as the detecting antibody was used in 117 HD patients. The lower limit of this assay was 0.1μg/ml, the average intra assay coefficient of variation (CV)was 2.2%, the inter assay CV was 9.0%, and the recovery rate was 97% ~107%. The median value of serum AGE was 0.5μg/ml in 117 HD patients, but only 9.4% of the age matched normal subjects had the serum AGE level above 0.1μg/ml. Compared with the results by using fluorescence spectrophotometry in 44 HD patients, analysis showed that the results by using ELISA was statistically correlated with those by using fluorescence spectrophtometry( P =0.01). These results indicate that competitive ELISA for measuring serum AGE is a simple, precise and specific method that can be used in the diagnosis of glycoxidative damage in HD patients.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2000年第6期391-393,共3页
Medical Journal of Chinese People's Liberation Army
基金
国家自然科学基金!资助课题 (编号 39770 349)
关键词
血液透析
肾功能衰竭
ELISA
糖基化终产物
Glycosylation end products, advanced
Enzyme linked immunoabsorbent assay
Hemodialysis
