磁免疫电化学发光高灵敏检测人血清中肿瘤标志物的研究

Tumor is a serious threat to human health. Tumor markers are an important index on diagnosis of cancer. The test for the treatment of cancer and prognosis is of great significance. As new analysis technology, immunomagnetic electrochemiluminescence is developed in recent years. It has both the sensitivity of electroluminescence and the high specificity of immunoassay. It is a novel immunoassay and is superior to radio immunoassay, enzyme immunoassay, fluorescent immunoassay and chemiluminescent immunoassay because of their excellent sensitivity, high selectivity, wide dynamic range, good stability and easy fabrication. Up to now, it is the best method in the solid phase immunoassays. Magnetic nanoparticles have attracted significant interest in recent years and have been widely studied for microbiology, biochemistry, bioanalytical chemistry, and biomedicine applications such as enhancement procedure, magnetic immobilization, bioelectrocatalysis, and drug targeting. The combination of magnetic and luminescence properties into a single micro- or nanocomposite system would be very useful in the biomedical and biopharmaceutical fields. A magnetic bead based ECL method with tris(2,2′-bipyridyl) ruthenium and tripropylamine reaction has been demonstrated to be a highly sensitive method for biorelated detection. Recently, the use of nanoparticles-based probes in biorelated applications has increased in popularity due to their unique physical and chemical properties. In this project, we shall assemble a new device of immunomagnetic electrochemiluminescence, synthesize superparamagnetic nanoparticles, prepare the magnetic beads of glutathione stransferase (GST), neuron specific endolase (NSE), carbohydrate antigen 153 (CA153), squamous cell carcinoma antigen （SCCA), cytokeratin fragment antigen 21-1 (CYFRA21-1) and tissue polypeptide specific antigen (TPS) and their antibody complexes. In addition, we shall also develop a series of novel methods for the determination of CYFRA21-1, NSE, SCCA, TPS, CA153 and GST by using immunomagnetic electrochemiluminescence and application to the detection of CYFRA21-1, NSE, SCCA, TPS, CA153 and GST in the serum of lung cancer patients. We shall establish the data base of corelation of tumer markers and lung cancer.

肿瘤是一类严重威胁人类健康的恶性疾病。肿瘤标志物作为诊断肿瘤的一项重要指标，其检测对于疾病治疗与预后具有重要意义。免疫电化学发光是近年发展起来的新型分析技术，它既有电化学发光检测的高度敏感性又有免疫分析的高度特异性，是继放射免疫、酶免疫、荧光免疫及化学发光免疫测定之后的新一代标记免疫测定技术，具有灵敏、快速和稳定的特点，在固相标记免疫测定中居领先地位。 本项目研制高灵敏度的磁免疫电化学发光仪；合成超顺磁性纳米磁球；制备非小细胞肺癌抗原(CYFRA21-1)、神经元特异性烯醇化酶(NSE)、鳞状细胞癌抗原(SCCA)、组织多肽特异性抗原(TPS)、糖类抗原(CA153)及谷光苷肽转换酶(GST)免疫纳米磁球及其抗体复合物；建立CYFRA21-1、NSE、SCCA、TPS、CA153及GST磁免疫电化学发光检测新方法，并用于检测肺癌患者血清中上述6种肿瘤标志物；建立肿瘤标志物与癌症相关性数据。

肿瘤是一类严重威胁人类生命和健康的恶性疾病，根据世界卫生组织(WHO)报道，全球范围内恶性肿瘤是人类仅次于心脑血管病的第二大死亡原因，占总死亡人数的22%，并逐年增加。因此，WHO指出早期诊断和治疗是对抗癌症的有效手段之一，肿瘤的早期诊断是决定其预后的一个重要因素。肿瘤标志物(Tumor marker，TM)作为一项重要指标，在肿瘤早期诊断当中发挥着越来越重要的作用。目前发现的肿瘤标志物已经有一百多种，由于大多数情况下体内肿瘤标志物的含量都非常低，一般的检测方法很难对其进行精确的定量。过去对肿瘤标志物的检测主要用放射免疫分析法，这种方法存在诸多缺点，如操作复杂、测定结果不稳定、试剂保存时间短、放射性污染、仪器昂贵等。因此，亟待发展一种新的检测方法对血液中TM 进行精确的定量分析。电化学发光法检测正好适应了这种需要。非小细胞肺癌抗原（CYFRA21-1）、神经元特异性烯醇化酶（NSE）、鳞状细胞癌抗原（SCCA）、组织多肽特异性抗原(TPS)、糖抗原(CA153)、 谷光苷肽转换酶(GST)是最有价值的肺癌患者血清肿瘤标志物，对早期诊断、疗效观察、预后监测有重要意义。该项目经过四年的研究，建立了CYFRA21-1、NSE、SCCA、TPS、CA153、GST免疫磁球抗原抗体复合物的电化学发光检测新方法，进行了检测肺癌患者血清中 CYFRA21-1，NSE，SCCA，TPS，CA153 及 GST 等肿瘤标志物。本项目共发表SCI论文15篇，其中SCI一区2篇、二区7篇，SCI影响因子3.0以上论文10篇，总SCI影响因子58.383（按2016年SCI期刊影响因子计），获得授权发明专利2项，参与英国皇家化学会出版著作一部，圆满地完成了项目研究内容和考核指标。上述肿瘤标志物磁免疫ECL分析方法的建立为肿瘤患者带来福音，为早期诊断肿瘤和人人享受健康具有重要的意义。本项目发表的论文被他人引用达40多次，其中铅氢化物发生机理研究受到国际著名原子光谱学家意大利国家研究院Alessandro D'Ulivo 教授专评，说明本项目资助的研究成果引起国际同仁的关注、产生一定的国际影响。