Planche Virginie, Brochet Christine, Bakkouch Asma, Bernard Maguy
Services de biochimie métabolique et de biochimie endocrinienne et oncologique, Groupe Hospitalier Pitié-Salpêtrière, APHP, Paris, France.
Ann Biol Clin (Paris). 2010 Mar-Apr;68(2):239-42. doi: 10.1684/abc.2010.0422.
The aim of this work is to establish a pre-analytical approach suitable for the neuron-specific enolase (NSE) measurement. This enzyme which is synthesized by neurons and neuroendocrine cells, is a marker useful for the diagnosis and the monitoring of patients with neuroendocrine tumors (neuroblastoma, small cell lung cancer) and during stroke to assess neuronal damage. This NSE measurement is very sensitive to hemolysis due to the abundance of the enzyme in red blood cells. Two methods of evaluation of hemolysis have been compared: the determination of free haemoglobin (Hb) by spectrophotometry and the indirect measurement of an hemolytic index with a multiparameter analyzer, the Modular (Roche Diagnostics). The correlation between these 2 methods on 42 samples is very satisfactory: Y (free Hb) = 12.337 X (index) + 31.743 r = 0.997. The NSE assay is based on TRACE (Time Resolved Amplified Cryptate Emission) technology, on a Kryptor (BRAHMS). The influence of hemolysis on the determination of NSE was confirmed by overloading with hemoglobin (hemolysate) 3 pools of serum with NSE concentrations close to the threeshold decision. The determination of NSE shows an increase in concentration parallely to the hemolytic index (about 150% for an hemolytic index of 10). Consequently, in our laboratory the NSE determination is realized only for samples presenting an hemolytic index < or = 10, this allowing a good monitoring of kinetics of this marker.
这项工作的目的是建立一种适用于神经元特异性烯醇化酶(NSE)检测的分析前方法。这种由神经元和神经内分泌细胞合成的酶,是用于诊断和监测神经内分泌肿瘤(神经母细胞瘤、小细胞肺癌)患者以及在中风期间评估神经元损伤的有用标志物。由于红细胞中该酶含量丰富,NSE检测对溶血非常敏感。比较了两种评估溶血的方法:通过分光光度法测定游离血红蛋白(Hb)以及使用多参数分析仪Modular(罗氏诊断)间接测量溶血指数。对42个样本的这两种方法之间的相关性非常令人满意:Y(游离Hb)= 12.337X(指数)+ 31.743,r = 0.997。NSE检测基于Kryptor(BRAHMS)上的TRACE(时间分辨放大镧系元素荧光免疫分析)技术。通过用血红蛋白(溶血产物)使3组NSE浓度接近临界值的血清过载,证实了溶血对NSE测定的影响。NSE测定显示浓度随溶血指数平行增加(溶血指数为10时约增加150%)。因此,在我们实验室,仅对溶血指数≤10的样本进行NSE测定,这样可以很好地监测该标志物的动力学变化。