预期应用
ELISA法定量测定大鼠血清中NSE含量，有助于诊断小细胞肺癌及相关性疾病。
概述
烯醇化酶（Enolase）是参与糖酵解的酶，催化2-磷酸甘油酸向磷酸烯醇丙酮酸的转化，由三种亚基（α、β、γ）组成的二聚体同功酶。其中ag 和 gg烯醇化酶同功酶存在于神经元及神经来源的细胞中，也称为神经元特异性烯醇化酶（Neurone specific enolase ，NSE）。在正常大鼠群中NSE含量较低，但在某些神经内皮细胞增生的恶性疾病，如神经母细胞瘤（Neuroblastoma），其含量往往上升。
由于小细胞肺癌（Small cell lung carcinoma，SCLC）是最常表现有神经内分泌性质的肿瘤，目前NSE也是SCLC最敏感最特异的肿瘤标志物。肺癌是近年来发病率日趋增高的癌种，在种种肺癌中小细胞肺癌占大约20%，因此正确诊断肺癌的类型，特别是确定小细胞肺癌的诊断很重要。小细胞肺癌患者的ag 和 gg同功酶含量均有不同比例的增高，因些必须在同一敏感度下同时检测NSE的ag和 gg同功酶含量。本试剂盒所用单抗只针对NSE的g亚基，而与a或β亚基无交差反应。
据文献报道，NSE含量测定可作为肺癌、成神经细胞瘤、黑素瘤、精原细胞瘤以及中枢精神系统损伤的诊断指标。同时，也可用于SCLC治疗前后疗效的监测与预后，以及SCLC与其它型肺癌的鉴别诊断。
实验原理
本试剂盒应用双抗体夹心酶标免疫分析法测定血清中NSE水平。用纯化的抗体包被微孔板，制成固相抗体，往包被单抗的微孔中依次加入NSE抗原、生物素化的抗大鼠NSE抗体、HRP标记的亲和素，经过彻底洗涤后用底物TMB显色。TMB在过氧化物酶的催化下转化成蓝色，并在酸的作用下转化成最终的黄色。颜色的深浅和样品中的NSE呈正相关。用酶标仪在450nm波长下测定吸光度（OD值），计算样品浓度。
Intended use
The Neuron-Specific Enolase (NSE) enzyme linked immunosorbent assay (ELISA) provides quantitative measurement of rat NSE in serum to aid in the clinical evaluation of patients suspected of having small cell lung cancer, and other related diseases.
Introduction
The glycolytic enzyme enolase (2-phosph-D-glycerate hydrolyase) exists as several dimeric isoenzymes (aa, ab, ag and gg) composed of three distinct subunits, a, b, and g. Three isoenzymes are found in rat brain: aa, ag and gg. The ag and gg-enolase isoenzymes are also known as neuron-specific enolase (NSE) as these isoenzymes initially were detected in neurons and neuronendocrine cells. The NSE levels are low in health and benign subjects. Elevated levels are commonly found in patients with malignant tumors with neuronendocrine differentiation, especially small cell lung cancer and neuroblasloma.
Lung cancer is one of the most spread cancer forms with incidences about 50~100 per 100,000 population. Approximately 20% of the lung cancer is small cell lung cancer. Patients with small cell lung cancer show various proportions of ag and gg isoenzyme. The determination of NSE should detect ag and gg isoforms with the same sensitivity . The antibodies for this particular assay are specific for the g-subunit without cross reactivity with a or b subunits.
NSE are reported to be useful diagnostic marker for lung cancer, neuroblastoma, melanoma, seminoma and in injury of central nervous system. In addition to the above, NSE can be a valuable tool in following-up the effect of chemotherapy of small cell lung cancer, in prognostic evaluation of patients with small cell lung cancer, and in differential diagnosis between cell lung cancer and non-small cell lung cancer.
Test principle
The NSE Quantitative Test Kit is based on a solid phase enzyme-linked immunosorbent assay. The assay system utilizes one monoclonal anti-gNSE antibody for solid phase (microtiter wells) immobilization and another biotinylated polyclonal anti-gNSE antibody in the antibody-enzyme conjugate solution. The standards and test specimen (serum) are added to the antibody coated microtiter wells. During the incubation, specific NSE bound to anti-NSE antibody on the wells. Unbound NSE antigen is removed by washing the wells with buffer. HRP labeled streptavidin
is then added to each well. After another incubation, unbound HRP labeled streptavidin is washed off and the amount of bound peroxidase is proportional to the concentration of the NSE present in each sample. Upon addition of the substrate and chromogen, the intensity of blue color will develop in proportion to the concentration of NSE antigen in the samples.
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