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

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

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.

Keywords: Acute myocardial infarction; Creatine kinase MB; Eu (III) chelate microparticles; Fluorescence immunochromatographic assay.

MeSH terms

  • Chelating Agents / chemistry*
  • Chromatography, Affinity / instrumentation
  • Chromatography, Affinity / methods*
  • Creatine Kinase, MB Form / metabolism*
  • Europium / chemistry*
  • Microspheres*
  • Polystyrenes / chemistry
  • Reagent Strips / chemistry
  • Reference Values
  • Spectrometry, Fluorescence
  • Time Factors

Substances

  • Chelating Agents
  • Polystyrenes
  • Reagent Strips
  • Europium
  • Creatine Kinase, MB Form