Simultaneous express immunoassay of multiple cardiac biomarkers with an automatic platform in human plasma
Natalia L Nechaeva1, Olga N Sorokina1, Tatyana S Konstantinova1
1N.M. Emanuel Institute of Biochemical Physics, Russian Academy of Sciences, 119334, Kosygina str. 4, Moscow, Russia.
Insights
This study introduces a rapid, magnetic bead-based method for simultaneously detecting four cardiac biomarkers: C-reactive protein, cystatin C, myoglobin, and D-dimer. This innovation aids in the early diagnosis of acute myocardial infarction and differential diagnostics.
Area of Science:
- Biochemistry
- Clinical Diagnostics
- Biotechnology
Background:
- C-reactive protein, cystatin C, myoglobin, and D-dimer are key indicators of inflammation and thromboembolic status.
- Early and differential diagnostics of acute myocardial infarction (AMI) rely on accurate assessment of these biomarkers.
- Current diagnostic methods may lack the speed and efficiency required for rapid clinical decision-making.
Purpose of the Study:
- To develop a rapid, simultaneous detection method for four critical cardiac biomarkers.
- To establish an express analysis technique using magnetic beads for improved diagnostic efficiency.
- To validate the method's applicability in human plasma samples for early AMI diagnostics.
Main Methods:
- Development of an immunomagnetic extraction technique for simultaneous analyte isolation from plasma.
- Utilizing fluorescent dye labeling for sensitive detection of biomarkers.
- Optimization of reaction times for quantification in spiked solutions and human plasma.
- Integration of single-protein detection into a four-protein analysis on an automated platform.
Main Results:
- A novel method enables the express analysis of C-reactive protein, cystatin C, myoglobin, and D-dimer.
- The technique requires a minimal sample volume (17 μL) and provides results within 14 minutes for all four proteins.
- The method demonstrates a wide dynamic range (over two orders of magnitude) and analytical applicability in human plasma.
- Successful measurements were performed on samples from 16 volunteers, confirming its practical utility.
Conclusions:
- The developed automated platform offers a highly efficient and rapid method for simultaneous quantification of four cardiac biomarkers.
- This technique significantly reduces analysis time and sample volume, crucial for emergency diagnostics like AMI.
- The method holds promise for improving early diagnosis and differential diagnosis of cardiovascular events.
Abstract:
C-reactive protein, cystatin C, myoglobin, and D-dimer represent the inflammatory or thromboembolic status of the patient and play important roles in early diagnostics of acute myocardial infarction. Each protein can indicate some health problems, but their simultaneous detection can be crucial for differential diagnostics. The express analysis of these proteins in a small drop of plasma was developed using magnetic beads. The suggested method is based on immunomagnetic extraction of the target analyte from plasma samples and its simultaneous labelling by fluorescent dye. Reaction time was optimized for quantification of cardiac biomarkers in the spike solutions and human plasma samples. In this paper, we developed a one-protein detection technique for each cardiac biomarker and united it to a four-protein facility using an automatic platform. The proposed technique requires only 17 μL of the human plasma and takes 14 min for four-protein measuring. The suggested technique covers concentration difference by more than two orders of magnitude and demonstrates analytical applicability by measurements of human plasma samples of 16 volunteers.
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