摘要
目的 建立尿中性肽内切酶 (NEP)检测的ELISA法 ,明确用ELISA法检测尿NEP在诊断肾小管损伤中的意义。方法 建立尿NEP检测的ELISA法 ,并与荧光法比较 ;分别用两法检测患者组和对照组尿NEP。结果 ELISA法检测NEP ,浓度在 2 0～ 2 0 0 μg/L时 ,浓度的对数与吸光度间有良好的线性关系 (r >0 .99,P <0 .0 1)。取 10次标准曲线 ,计算线性范围内各浓度的变异系数 (CV)为 2 .0 6 %～ 4 .17% ,平均 2 .98%。所有标准曲线相关系数 (r)均 >0 .99(P <0 .0 0 1)。ELISA法回收率为 93.6 %～ 10 7.3% ,平均为 10 1.7%。该法批内CV为 4 .15 %、5 .4 6 % ,批间CV为 6 .2 4 %、6 .39%。检测限值为 2 .2 0 μg/L。患者组两法测得尿NEP均低于正常对照组 (P <0 .0 1)。各组两法结果无差异 (P均 >0 .0 5 )。结论 检测尿NEP可以帮助判断近端肾小管损伤。ELISA法与传统荧光法相比更简便 ,不需特殊仪器 ,却有良好的精确度和重复性 ,可以作为评价肾小管损伤的较好方法。
Objective To set up an ELISA method to measure urinary neutral endopeptidase(NEP) and determine its clinical meaning in diagnosing renal tubular injury. Methods Set up an ELISA method and compare it with fluorimetric assay. Determine urinary NEP of both patient group and control group by both methods. Results In ELISA when the lever of urinary NEP was within 20 to 200 μg/L, the logarithm of its lever and its A value had proper linear relationship( r >0.99, P <0.01). In the ten standard curves, the CV of all the concentration within its linear range was 2.06%～4.17% and mean was 2.98%, r value of each standard curve was over 0.99 ( P <0. 001 ). The recovery rate was 93.6%～107.3%, and the mean was 101.7%. The difference within batch were 4.15% , 5.46%, and the difference between batch were 6.24%, 6.39% respectively. The least value of this method was 2.20 μg/L. The urinary NEP level of patient group measured by both methods was much lower than that of control group and the difference of the two methods in each group had no significance. Conclusions Measurement of urinary NEP is helpful in diagnosing renal tubular injury. ELISA is easier to use than fluorimetric assay, and a fluorimetric apparatus is not essential. But the precision and reproducibility were good. So it can be use to evaluate renal tubular injury.
出处
《上海医学检验杂志》
北大核心
2003年第5期281-284,共4页
Shanghai Journal of Medical Laboratory Sciences
基金
上海市卫生局医学领先专业学科基金 ( 983 0 0 9)
