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Related Experiment Video

Updated: May 22, 2026

Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction
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An aptamer-capture based chromogenic assay for thrombin.

Qiang Zhao1, Xiaofang Wang

  • 1Research Center for Environmental Science and Engineering, Shanxi University, Taiyuan, China. chemzhaoq@hotmail.com

Biosensors & Bioelectronics
|March 6, 2012
PubMed
Summary

A novel aptamer-based assay effectively captures and quantifies human alpha thrombin using magnetic beads and a chromogenic reaction. This sensitive and specific method achieves detection limits as low as 40 fM, suitable for serum analysis.

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Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Biotechnology

Background:

  • Accurate quantification of human alpha thrombin is crucial for diagnosing and monitoring various hemostatic and thrombotic disorders.
  • Existing methods for thrombin detection may lack the required sensitivity, specificity, or speed for certain clinical applications.
  • Aptamer-based assays offer a promising alternative due to their high affinity and specificity for target molecules.

Purpose of the Study:

  • To develop a simple, sensitive, and specific chromogenic assay for the detection and quantification of human alpha thrombin.
  • To utilize aptamer affinity capture combined with enzymatic amplification for enhanced detection capabilities.
  • To validate the assay's performance, including sensitivity, specificity, and applicability to biological samples like human serum.

Main Methods:

  • Human alpha thrombin was captured using aptamer-modified magnetic beads, facilitating sample enrichment.
  • A subsequent enzymatic reaction catalyzed the conversion of a chromogenic substrate by the captured thrombin.
  • The generated colored product was quantified using an absorbance spectrometer to determine thrombin concentration.

Main Results:

  • The assay demonstrated high sensitivity, achieving a concentration detection limit of 40 fM with a 24-hour enzyme reaction using tosyl-Gly-Pro-Arg-p-nitroanilide substrate.
  • A rapid analysis option enabled detection of 400 fM thrombin within a 2-hour enzyme reaction period.
  • The assay exhibited good specificity, attributed to selective aptamer binding and the specific enzymatic reaction, and was successfully applied to detect thrombin in dilute human serum.

Conclusions:

  • A simple, sensitive, and specific aptamer-based chromogenic assay for human alpha thrombin has been successfully developed.
  • The assay leverages aptamer affinity capture and enzymatic amplification, offering low detection limits and applicability to serum samples.
  • This method provides a valuable tool for rapid and accurate thrombin quantification in clinical and research settings.