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

肖春海，梁爽，刘静凡，李飞飞，朱庆华，瞿培培，董志武

(上海市第六人民医院金山分院检验科，上海 201500)

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

Evaluation of BNP quantification by single-epitope sandwich combined with cyclic enhanced immunofluorescent assayXIAOChunhai，LIANGShuang，LIUJingfan，LIFeifei，ZHUQinghua，QUPeipei，DONGZhiwu

.(DepartmentofClinicalLaboratory，JinshanBranchofShanghaiSixthPeople′sHospital，Shanghai201500，China)

Abstract:ObjectiveTo evaluate B-type natriuretic peptide quantification by single-epitope sandwich(SES)combined with cyclic enhanced immunofluorescent assay(CEIFA). MethodsBy 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). ResultsThe 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. ConclusionsSES 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

B型钠尿肽(B-type natriuretic peptide，BNP)属于利钠肽家族，由日本学者首先从猪脑中分离出来，但却主要由心室肌细胞分泌，与左心室承受的压力大小密切相关，所以现在更多称“B型利钠肽”。人类BNP基因位于1号染色体端臂远端，包括2个内含子和3个外显子。BNP前体(BNP precursor, proBNP)裂解后同时产生具有环状结构的BNP和氨基末端碎片(N-terminal proBNP，NT-proBNP)，两者都可作为心力衰竭诊断的有效特异指标[1-3]。BNP虽然在诊断呼吸困难、肺栓塞、肺癌等疾病的过程中发挥重要作用[4-6]，但目前应用较多的仍为心血管领域[1-3，7-10]。我们主要是利用循环增强荧光免疫技术(cyclic enhanced immunofluorescent assay, CEIFA)结合单一位点夹心抗体(single-epitope sandwich，SES)法[11]实现BNP的检测。

材料和方法

一、材料

1. 研究对象45份上海市第六人民医院金山分院心内科患者的乙二胺四乙酸二钾盐新鲜抗凝血浆；星童公司BNP标准品，浓度为100、1 000和5 000 pg/mL。……