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Aptamer-based fluorometric lateral flow assay for creatine kinase MB.

Jing Zhang1, Xuefei Lv1, Wei Feng1

  • 1Beijing Key Laboratory for Separation and Analysis in Biomedicine and Pharmaceuticals, School of Life Science, Beijing Institute of Technology, 5 South Zhongguancun Street, Haidian District, Beijing, 100081, People's Republic of China.

Mikrochimica Acta
|July 9, 2018
PubMed
Summary

Researchers developed a sensitive and specific aptamer-based lateral flow assay for detecting creatine kinase MB (CKMB). This portable test offers accurate results for CKMB quantification, aiding in potential cardiac marker diagnostics.

Keywords:
Acute myocardial infarctionAptamerCKMBFluorescent microsphereLateral flowMagnetic SELEXNanoparticlesRapid detectionSandwichTest strip reader

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Area of Science:

  • Biotechnology
  • Biomedical Diagnostics
  • Analytical Chemistry

Background:

  • Creatine kinase MB (CKMB) is a crucial biomarker for acute myocardial infarction (AMI).
  • Existing diagnostic methods for CKMB can be complex or lack portability.
  • Development of rapid, sensitive, and specific detection methods is essential for early AMI diagnosis.

Purpose of the Study:

  • To develop and validate a novel aptamer-based lateral flow assay for sensitive and specific detection of CKMB.
  • To evaluate the performance characteristics of the assay, including limit of detection, dynamic range, and specificity.
  • To assess the assay's accuracy using spiked serum samples.

Main Methods:

  • Selection of high-affinity aptamers for CKMB using magnetic systematic evolution of ligands by exponential enrichment.
  • Development of a fluorescent microsphere-based lateral flow assay incorporating selected aptamers (C.Apt.21 and C.Apt.30).
  • Characterization of assay performance, including limit of detection, concentration range, specificity against other biomarkers, and accuracy in serum samples.

Main Results:

  • Two aptamers, C.Apt.21 and C.Apt.30, demonstrated high affinity and specificity for CKMB.
  • The developed aptamer-based lateral flow assay is portable, economical, and sensitive, with a limit of detection of 0.63 ng·mL⁻¹.
  • The assay exhibits a wide working concentration range (0.005 - 2 μg·mL⁻¹), specificity for CKMB, and high accuracy (88-117% recovery) in spiked serum samples, with no interference from cardiac troponin I or myoglobin.

Conclusions:

  • The developed aptamer-based lateral flow assay provides a sensitive, specific, and accurate method for CKMB detection.
  • This assay holds promise as a point-of-care diagnostic tool for AMI.
  • The use of selected aptamers in a lateral flow format offers a viable alternative to traditional CKMB detection methods.