ELISA双抗体夹心法检测嗜铬蛋白A的性能评价及临床应用  

Performance of double-antibody sandwich ELISA for chromogranin A determination

作　　者：胡玉懿 吴蕙 韩序 张春燕 郭玮 楼文晖 潘柏申 

机构地区：[1]复旦大学附属中山医院检验科,上海200032 [2]复旦大学附属中山医院普外科,上海200032

出　　处：《检验医学》2018年第1期55-59,共5页Laboratory Medicine

基　　金：国家自然科学基金面上项目(81572064);上海市卫生计生系统重要薄弱学科建设项目(2015ZB0201);上海市卫生系统先进适宜推广技术项目(2013SY065);上海申康医院发展中心课题(SHDC22014017)

摘　　要：目的验证酶联免疫吸附试验(ELISA)双抗体夹心法检测嗜铬蛋白A(Cg A)的性能,评价其初步的临床应用价值。方法方法学验证参考美国临床实验室标准化协会(CLSI)EP15-A、EP6-A和EP17-A文件,对项目的精密度、线性和定量检测下限进行验证。采用厂商提供的质控品验证正确度。检测66例神经内分泌肿瘤患者、41名健康对照者和65例疾病对照患者的Cg A水平,采用受试者工作特征(ROC)曲线评估Cg A对神经内分泌肿瘤患者的诊断能力。结果 ELISA双抗体夹心法检测血清Cg A的批内变异系数(CV)<14.00%、批间CV<8.00%;正确度验证测定值的均值在标示浓度范围内,相对偏移为4.51%;在8~740 ng/m L的范围内线性良好(Y=0.793X-3.791,r2=0.988);定量检测下限为7.93 ng/m L。神经内分泌肿瘤组、健康对照组和疾病对照组之间血清Cg A水平差异均有统计学意义(P<0.000 1);Cg A鉴别正常人与神经内分泌肿瘤患者的ROC曲线下面积(AUC)为0.741,最佳诊断界值为47.18 ng/m L,诊断敏感性为78.8%,特异性为68.3%。结论 ELISA双抗体夹心法检测Cg A试剂盒具有良好的精密度、正确度、线性范围和定量检测下限,检测性能符合临床要求。血清Cg A水平在神经内分泌肿瘤患者中显著升高,因此可用于鉴别健康人和神经内分泌肿瘤患者。Objective To evaluate the performance of double-antibody sandwich enzyme-linked immunosorbent assay （ELISA）for chromogranin A （CgA）determination. Methods The precision,linearity and limit of determination of CgA determination by double-antibody sandwich ELISA were verified according to the Clinical and Laboratory Standards Institute （CLSI） EP15-A,EP6-A and EP17-A. The accuracy of CgA determination was assessed by manufacturer＇s quality control materials. A total of 66 neuroendocrine neoplasm patients,41 healthy subjects and 65 disease control patients were enrolled. The CgA levels in the 3 groups were determined,and receiver operating characteristic （ROC） curve was used to compare the efficiency of CgA determination in diagnosing neuroendocrine neoplasm. Results The within-run and between-run coefficients of variation （CV） of CgA determination by double-antibody sandwich ELISA were 〈14.00% and 〈8.00%,respectively. For accuracy evaluation,the relative bias was 4.51%,when the mean of results was within the indicated range. There was a good linear correlation（Y=0.793X-3.791,r2=0.988） in the range of 8-740 ng/mL. The limit of determination was 7.93 ng/mL. The difference of CgA levels in neuroendocrine neoplasm,healthy control and disease control groups had statistical significance （P〈0.000 1）. The area under curve（AUC）of CgA determination in distinguishing healthy subjects and neuroendocrine neoplasm patients was 0.741,when the cut-off value was 47.18 ng/mL. The sensitivitywas 78.8%,and the specificity was 68.3%. Conclusions CgA determination by double-antibody sandwich ELISA has good performance in precision,accuracy,linearity and limit of determination,which meets clinical needs. Serum CgA levels increase in patients with neuroendocrine neoplasm. CgA could be a marker for distinguishing healthy subjects and neuroendocrine neoplasm patients.

关 键 词：嗜铬蛋白A 神经内分泌肿瘤 敏感性 特异性 受试者工作特征曲线 曲线下面积 

分 类 号：R446.61[医药卫生—诊断学]