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一种使用铕(III)螯合物微粒的荧光免疫层析测定法,用于快速、定量和灵敏地检测肌酸激酶同工酶MB。

A Fluorescence Immunochromatographic Assay Using Europium (III) Chelate Microparticles for Rapid, Quantitative and Sensitive Detection of Creatine Kinase MB.

作者信息

Lai Xiao-Hong, Liang Rong-Liang, Liu Tian-Cai, Dong Zhi-Ning, Wu Ying-Song, Li Lin-Hai

机构信息

Department of Laboratory Medicine, Guangzhou General Hospital of Guangzhou Military Command of PLA, No.111, Liuhua Road, Yuexiu District, Guangzhou, Guangdong, China.

Guangzhou University of Chinese Medicine, No.12, Airport Road, Baiyun District, Guangzhou, Guangdong, China.

出版信息

J Fluoresc. 2016 May;26(3):987-96. doi: 10.1007/s10895-016-1786-3. Epub 2016 Apr 1.

DOI:10.1007/s10895-016-1786-3
PMID:27034063
Abstract

The isoenzyme creatine kinase MB is very important for diagnosis of acute myocardial infarction (AMI). Some CK-MB immunoassays are sensitive, accurate and available for clinical application, but they are expensive and time-consuming procedures. Furthermore, conventional fluorescence immunochromatographic assays (FL-ICAs) have suffered from background fluorescence interference and low analytical sensitivity. A rapid and simple FL-ICA with Eu (III) chelate polystyrene microparticles was developed to determine CK-MB in 50uL serum samples using a portable test strip reader by measuring the fluorescence peak heights of the test line (HT) and the control line (HC) in 12 min. The assay was reliable with a good correlation coefficient between HT/HC ratio and CK-MB concentration in samples. A linear range was 0.85-100.29 ng/mL for CK-MB, and the LOD was 0.029 ng/mL. The intra- and inter-assay coefficients of variation (CV) were both <10 % and the average recoveries were from 90.17 % -112.63 % for CK-MB. The system performed well in interference experiments. Furthermore, a highly significant correlation (r = 0.9794, P < 0.001) between this method and the commercially available bioMérieux mini VIDAS system were attained for measuring 120 CK-MB samples. These results indicated that the Eu (III) chelate microparticles-based FL-ICA is simple, fast, highly sensitive, reliable, and reproducible for point-of-care testing of CK-MB concentrations in serum. Graphical Abstract ᅟ.

摘要

肌酸激酶同工酶MB对急性心肌梗死(AMI)的诊断非常重要。一些CK-MB免疫测定法灵敏、准确且可用于临床应用,但它们成本高昂且耗时。此外,传统的荧光免疫层析测定法(FL-ICA)存在背景荧光干扰和分析灵敏度低的问题。我们开发了一种基于铕(III)螯合聚苯乙烯微粒的快速简便的FL-ICA,通过使用便携式试纸条读数器在12分钟内测量测试线(HT)和对照线(HC)的荧光峰高,来测定50μL血清样本中的CK-MB。该测定法可靠,样品中HT/HC比值与CK-MB浓度之间具有良好的相关系数。CK-MB的线性范围为0.85 - 100.29 ng/mL,检测限为0.029 ng/mL。CK-MB的批内和批间变异系数(CV)均<10%,平均回收率为90.17% - 112.63%。该系统在干扰实验中表现良好。此外,对于120个CK-MB样本的测量,该方法与市售的生物梅里埃mini VIDAS系统之间具有高度显著的相关性(r = 0.9794,P < 0.001)。这些结果表明,基于铕(III)螯合微粒的FL-ICA对于血清中CK-MB浓度的即时检测简单、快速、高度灵敏、可靠且可重复。图形摘要ᅟ。

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