单一位点夹心抗体法结合循环增强荧光免疫技术检测BNP的评估

doi:10.3969/j.issn.1673-8640.2015.05.020

摘要/Abstract

摘要： 目的初步评价单一位点夹心抗体(SES)法结合循环增强荧光免疫技术(CEIFA)检测B型钠尿肽(BNP)的性能。方法用SES法结合CEIFA仪检测BNP,对其精密度、线性范围、功能敏感性、回收率、样本稳定性及与西门子ADVIA Centaur CP化学发光仪(简称Centaur CP)的相关性进行评估。结果 SES法结合CEIFA的批内和批间变异系数(CV)分别为4.38%~7.84%和10.47%~11.21%。线性范围为25~5 000 pg/mL。功能敏感性为12.5 pg/mL。回收率为95.04%~109.2%。与Centaur CP比较,其回归方程为Y=0.998 3X-22.38(r=0.969,P<0.01)。对样本稳定性的要求低于Centaur CP。结论 SES法结合CEIFA测定BNP具有较好的性能,与传统的双抗体夹心发光法比较有一定优势,适合临床急诊实验室使用。

关键词: B型钠尿肽, 单一位点夹心抗体, 荧光免疫分析, 心力衰竭

Abstract: Objective To evaluate B-type natriuretic peptide quantification by single-epitope sandwich(SES)combined with cyclic enhanced immunofluorescent assay(CEIFA). Methods By SES combined with CEIFA, BNP was determined. The precision, linear range, function sensitivity, recovery rate and sample stability were evaluated, and 45 samples were determined by the method, which the results were evaluated with those of SIEMENS ADVIA Centaur CP chemiluminescence analyzer (Centaur CP). Results The within-run and between-run coefficients of variation(CV) were 4.38%-7.84% and 10.47%-11.21%, respectively. The linear range was 25-5 000 pg/mL. The function sensitivity was 12.5 pg/mL. The recovery rate was 95.04%-109.2%. The correlation formula between the novel method and Centaur CP was Y=0.998 3X-22.38 (r=0.969, P<0.01). The request of sample stability of SES combined with CEIFA was lower than that of Centaur CP. Conclusions SES combined with CEIFA for BNP quantification has a good performance, and there are some advantages compared with traditional double-antibody sandwich chemiluminescence assay.

Key words: B-type natriuretic peptide, Single-epitope sandwich, Immunofluorescent assay, Heart failure

中图分类号:

R446.61

肖春海, 梁爽, 刘静凡, 李飞飞, 朱庆华, 瞿培培, 董志武. 单一位点夹心抗体法结合循环增强荧光免疫技术检测BNP的评估[J]. 检验医学, 2015, 30(5): 495-499.

XIAO Chunhai, LIANG Shuang, LIU Jingfan, LI Feifei, ZHU Qinghua, QU Peipei, DONG Zhiwu. Evaluation of BNP quantification by single-epitope sandwich combined with cyclic enhanced immunofluorescent assay[J]. Laboratory Medicine, 2015, 30(5): 495-499.

图/表 8

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样本	理论值 (pg/mL)	第1次 (pg/mL)	第2次 (pg/mL)	第3次 (pg/mL)	平均值 (pg/mL)	相对偏差 (%)
高值标准物不稀释	5 000	4 840	4 848	4 895	4 861.0	2.78
高值标准物2倍稀释	2 500	2 412	2 358	2 369	2 379.7	4.81
中值标准物不稀释	1 000	981	976	977	978.0	2.20
中值标准物2倍稀释	500	519	521	529	523.0	-4.60
中值标准物4倍稀释	250	241	235	239	238.3	4.67
低值标准物不稀释	100	89	91	95	91.7	8.33
低值标准物2倍稀释	50	54	49	53	52.0	-4.00
低值标准物4倍稀释	25	27	25	30	27.3	-9.33
低值标准物8倍稀释	12.5	17	18	11	15.3	-22.67

样本	理论值 (pg/mL)	第1次 (pg/mL)	第2次 (pg/mL)	第3次 (pg/mL)	平均值 (pg/mL)	相对偏差 (%)
高值标准物不稀释	5 000	4 840	4 848	4 895	4 861.0	2.78
高值标准物2倍稀释	2 500	2 412	2 358	2 369	2 379.7	4.81
中值标准物不稀释	1 000	981	976	977	978.0	2.20
中值标准物2倍稀释	500	519	521	529	523.0	-4.60
中值标准物4倍稀释	250	241	235	239	238.3	4.67
低值标准物不稀释	100	89	91	95	91.7	8.33
低值标准物2倍稀释	50	54	49	53	52.0	-4.00
低值标准物4倍稀释	25	27	25	30	27.3	-9.33
低值标准物8倍稀释	12.5	17	18	11	15.3	-22.67

标准液浓度 (pg/mL)	加入标准液前样本浓度 (pg/mL)	加入标准液后样本浓度 (pg/mL)	实际回收质量 (pg)	实际加入质量 (pg)	回收率 (%)
100	258	220.8	5.46	5	109.20
2 500	258	787.5	118.80	125	95.04

标准液浓度 (pg/mL)	加入标准液前样本浓度 (pg/mL)	加入标准液后样本浓度 (pg/mL)	实际回收质量 (pg)	实际加入质量 (pg)	回收率 (%)
100	258	220.8	5.46	5	109.20
2 500	258	787.5	118.80	125	95.04

时间 (h)	1号样本		2号样本		3号样本
时间 (h)	Pylon Core	Centaur CP	Pylon Core	Centaur CP	Pylon Core	Centaur CP
0	1 739	1 709	421	415	95	93
2	1 689	1 538	401	355	93	92
4	1 659	1 451	395	365	92	91
6	1 669	1 369	395	310	95	70
8	1 489	1 187	320	235	75	62
均值	1 649	1 451^*	386^*	336	90	82