ELISA双抗体夹心法检测嗜铬蛋白A的性能评价及临床应用

doi:10.3969/j.issn.1673-8640.2018.01.010

摘要/Abstract

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（CgA）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CgA水平,采用受试者工作特征（ROC）曲线评估CgA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CgA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/mL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/mL。神经内分泌肿瘤组、健康对照组和疾病对照组之间血清CgA水平差异均有统计学意义（P<0.000 1）;CgA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/mL,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CgA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CgA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。

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

Abstract:

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 o. 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 sensitivity was 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.

Key words: Chromogranin A, Neuroendocrine neoplasm, Sensitivity, Specificity, Receiver operating characteristic curve, Area under curve

中图分类号:

R446.61

胡玉懿, 吴蕙, 韩序, 张春燕, 郭玮, 楼文晖, 潘柏申. ELISA双抗体夹心法检测嗜铬蛋白A的性能评价及临床应用[J]. 检验医学, 2018, 33(1): 55-59.

HU Yuyi, WU Hui, HAN Xu, ZHANG Chunyan, GUO Wei, LOU Wenhui, PAN Baishen. Performance of double-antibody sandwich ELISA for chromogranin A determination[J]. Laboratory Medicine, 2018, 33(1): 55-59.

图/表 4

参考文献 24

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样本	批内		批间
样本	检测值	CV（%）	检测值	CV（%）
S1	41.70±5.49	13.17	41.98±3.08	7.33
S2	88.06±3.86	4.38	80.39±3.75	4.66
S3	298.04±15.00	5.03	294.88±10.60	3.93

样本	批内		批间
样本	检测值	CV（%）	检测值	CV（%）
S1	41.70±5.49	13.17	41.98±3.08	7.33
S2	88.06±3.86	4.38	80.39±3.75	4.66
S3	298.04±15.00	5.03	294.88±10.60	3.93

组别	例数	性别（男/女）	年龄（岁）	CgA（ng/mL）
胰腺神经内分泌肿瘤组	39	15/24	54.38±12.81	62.41（47.44,125.86）
胃神经内分泌肿瘤组	10	4/6	55.40±7.49	131.57（96.73,520.09）
肠神经内分泌肿瘤组	15	5/10	59.33±10.81	71.13（36.26,94.04）
肺神经内分泌肿瘤组	2	1/1	61/70^*	45.99/870.00^#

组别	例数	性别（男/女）	年龄（岁）	CgA（ng/mL）
胰腺神经内分泌肿瘤组	39	15/24	54.38±12.81	62.41（47.44,125.86）
胃神经内分泌肿瘤组	10	4/6	55.40±7.49	131.57（96.73,520.09）
肠神经内分泌肿瘤组	15	5/10	59.33±10.81	71.13（36.26,94.04）
肺神经内分泌肿瘤组	2	1/1	61/70^*	45.99/870.00^#