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Development of a Quantitative Detection Card for Heart-type Fatty Acid-binding Protein based on Background
Wei Zhang1, Junlei Chen2, Xinxia Li1,3
1Pharmacy Academy of Xinjiang Medical University, Urumqi, 830054, China.
Insights
A new, fast, and sensitive fluorescence quenching immunoassay was developed for heart-type fatty acid-binding protein (H-FABP) detection in serum. This method offers accurate H-FABP quantification for broader clinical applications.
Area of Science:
- Biochemistry
- Immunology
- Analytical Chemistry
Background:
- Heart-type fatty acid-binding protein (H-FABP) is a biomarker for cardiac injury.
- Accurate and rapid detection of H-FABP in serum is crucial for clinical diagnosis.
- Existing detection methods may lack speed or simplicity.
Purpose of the Study:
- To develop a fast, simple, and quantitative method for H-FABP detection in serum.
- To utilize a background fluorescence quenching immunochromatographic assay for enhanced sensitivity.
Main Methods:
- Developed a quantitative detection card using a double-antibody sandwich method with background fluorescence quenching.
- Optimized concentrations of coating and gold-labeled antibodies.
- Validated the method using clinical serum samples and common interfering substances.
Main Results:
- Achieved a sensitivity of 1.15 ng/mL with a linear range of 0-100 ng/mL.
- Demonstrated high specificity, with no significant cross-reactivity to common interfering substances.
- Clinical validation showed good correlation (R=0.95) with standard methods.
Conclusions:
- The developed assay is specific and sensitive for H-FABP detection in serum.
- This novel method has the potential for widespread application in clinical settings.
- Offers a rapid and reliable tool for H-FABP quantification.
Background:
To establish a fast and simple quantitative method for detection of heart-type fatty acid-binding protein (H-FABP) in serum based on a background fluorescence quenching immunochromatographic assay.
Methods:
A detection card based on the double-antibody sandwich double-antibody method with background fluorescence quenching was developed for quantitative measurement of H-FABP in serum. The optimal concentrations of control for coating the test and control lines were determined as well as the concentrations of gold-labeled antibodies used in preparing the detection system. The detection method for H-FABP in serum was established and validated using real-world clinical samples.
Results:
The optimal concentrations of labeling antibody and coating antibody were 5.0 μg/mL and 1.0 mg/mL, respectively. The test card had a sensitivity of 1.15 ng/mL over a linear concentration range of 0-100 ng/mL. Based on three batches prepared for testing the card, the relative standard deviation (RSD) within batches was less than 15% without a significant difference (P=0.942). The detection method was tested against common interfering substances in serum, such as bilirubin, triglyceride and serum anticoagulants ethylenediamine tetraacetic acid (EDTA), heparin, and sodium citrate, and no significant cross-reaction was detected. The test method was further validated with 50 clinical serum samples, and the test results were comparable with standard reference detection methods with good correlation (R=0.95).
Conclusion:
Our study presents a new method with strong specificity and sensitivity for the detection of H-FABP in serum, which could promote H-FABP detection in a broad range of applications.
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