摘要
[目的]建立双抗体夹心法检测血浆中热休克蛋白70(heatshockprotein70,HSP70)水平。[方法]制备兔抗HSP70抗体 ,建立双抗体夹心法标准曲线 ,鉴定其最小检出限、精确度、回收率 ,并初步应用于检测40例男性健康成人血浆样本的HSP70水平。[结果]双抗夹心ELISA法检测HSP70的最小检出限为10ng/ml,标准曲线在10～100ng/ml范围内线性良好。3份同批血清标本分别重复8次测定 ,平均批内变异系数为7.6 % ;3份不同批血清分别重复6次测定 ,平均批间变异系数为12.7 %。血清中加入已知量的标准抗原 ,测得回收率为85.5 %。对40例男性健康成人(20.0±0.9)岁血浆样本检测 ,其HSP70浓度均值为 (151.0±13.8)ng/ml。[结论]本方法检测HSP70的可测范围广 ,灵敏度和精密度较高变异系数较小 ,表明其灵敏、特异。
ObjectiveTo develop double antibodies sandwich-ELISA for detecting HSP70in plasma.[Methods] We established the method by using rabbit anti-sera against HSP70,and the lowest limit of detection,precision,recovery rate were tested respectively and40plasma in healthy males were examined using this method.[Results]The lowest limit of detection is10ng/ml;the linearity of the standard curve is preferable from10to100ng/ml.When three sera plasma samples were assayed repeatedly for8times and other3samples from different batch were assayed for6times,the average intra-assay and the inter-assay CV were7.6%and12.7%respecˉtively.The recovery rate was85.5%when some quantitative standard antigens were added into the plasma.The average concentrations of HSP70in plasma of40healthy males,20.0±0.9years old of age,were151±13.8ng/ml.[Conclusion]The established method for detection of HSP70will be useful because of its wide measure range,fine sensitivity,precision and a lowcoefficient of variation,and this method is sensitive,specific and stable.
出处
《环境与职业医学》
CAS
北大核心
2004年第1期27-30,共4页
Journal of Environmental and Occupational Medicine
基金
武汉科委晨光计划资助 (编号 :995004085)
