心肌损伤多肽和蛋白类标志物方法学分析和进展

doi:10.3969/j.issn.1673-8640.2023.08.015

摘要/Abstract

摘要：

国内外心肌损伤类疾病的患病率逐年升高，开展预防性监测，准确、及时的诊断是降低疾病危害的重要方法。目前，已知与心肌损伤密切相关的生物标志物绝大部分为多肽和蛋白类物质，其在外周血中的水平一般低于10^-8 g/L。基于免疫原理的检测技术可以对低丰度目标物开展检测，并已发展成为主流检测方法，但某些项目在检测准确度上仍存在固有问题。文章以B型钠尿肽（BNP）、肌钙蛋白等心肌损伤指标为例，对检验方法中影响结果准确度的典型问题进行分析，同时对新兴分析技术及其在心肌损伤标志物检测中的应用前景进行阐述。

关键词: 生物标志物, 心肌损伤, 临床检测方法, 新兴技术

Abstract:

The prevalence of myocardial injury diseases is increasing rapidly in the world. Monitoring and accurate diagnosis of myocardial injury diseases is essential to reduce disease hazard. Currently，the biomarkers in myocardial injury mainly belong to peptides and proteins，typically expressed lower than 10^-8 g/L. Immunoassay-based techniques enable a sensitive determination level for low abundant biomarkers，followed by developed as the dominant method. However，some inherent problems have also existed among current clinical immunoassays regarding testing accuracy. To illustrate testing barriers，this review proposes and summarizes some representative issues that usually interfere with the testing accuracy in B-type natriuretic peptide（BNP） and troponin. Meanwhile，some emerging technologies will be introduced in the context，and then to discuss its application that potential applied in myocardial injury clinical measurement.

Key words: Biomarker, Myocardial injury, Clinical determination, Emerging technology

中图分类号:

R446.1

孙泽朋, 王洪彬, 王建东, 宋德伟, 肖鹏. 心肌损伤多肽和蛋白类标志物方法学分析和进展[J]. 检验医学, 2023, 38(8): 784-789.

SUN Zepeng, WANG Hongbin, WANG Jiandong, SONG Dewei, XIAO Peng. Analysis and progress of peptide and protein biomarker methodology for myocardial injury[J]. Laboratory Medicine, 2023, 38(8): 784-789.

图/表 3

图1 基于SERS的夹心免疫分析法[14] 注：（a）用1层薄而光滑的金在玻璃载玻片上制成的捕获基板，抗体附着在金薄层表面；（b）抗体从样本中捕获蛋白标志物，加入抗体或适体（RNA，单链或双链DNA，可与靶分子特异性结合，其作用与抗原-抗体结合相似）和拉曼报告分子修饰的金纳米颗粒，作为SERS探针与蛋白标志物形成夹心结构；（c）激光激发附着在金纳米粒子上的拉曼报告分子产生拉曼光谱；（d）SERS信号被检测，其响应强度与结合的被测物的数量有关。

图2 悬浮阵列技术检测AMI生物标志物[17] 注：（a）羧基荧光编码微球；（b）荧光编码微球偶联AMI的生物标志物；（c）抗体与在荧光编码微球上的AMI生物标志物结合；（d）结合生物素化的第2抗体；（e）偶联R-藻红蛋白标记链霉亲和素的荧光报告分子（SA-PE）与第2抗体进行偶联，并获得平均荧光强度值；微球，生物标志物，第1抗体，生物素化第2抗体，R-藻红蛋白标记链霉亲和素的荧光报告分子。

图3 单分子阵列免疫分析示意图[18] 注：（a）蛋白分子首先被捕获在有抗体修饰磁珠上，用生物素化检测抗体和链霉亲和素-β半乳糖苷酶标记蛋白；（b）过量的磁珠被添加到含有低浓度蛋白的样本中，每个磁珠上只有1个或0个蛋白分子结合；将磁珠复合物装载到fL大小的微孔中，每个微孔只能容纳1个磁珠；用油密封阵列，获得阵列的荧光图像；磁珠，捕获抗体，抗原，生物素化检测抗体，链霉亲和素-β半乳糖苷酶。

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