研究目的
Lowering the limit of detection and enhancing the signal quality (color intensity) in lateral flow immunochromatographic assays through simple geometrical modifications.
研究成果
Geometrical modifications, specifically narrowing the detection pad width and optimizing test line position, significantly enhance the performance of lateral flow immunochromatographic assays by lowering the detection limit to 5 IU/L and increasing color intensity by up to 400%. These changes improve flow velocity control and analyte to antibody ratio, offering a cost-effective alternative to complex enhancement methods.
研究不足
The study is limited to hCG as the analyte; results may vary with other analytes. The method relies on visual and RGB-based quantification, which might have sensitivity constraints. The trapezoidal geometry modifications may not be universally applicable to all LFIA designs.
1:Experimental Design and Method Selection:
The study involved designing lateral flow immunochromatographic assay strips with varying detection pad widths (trapezoidal shapes) and test line positions to investigate their effects on flow velocity, analyte to antibody ratio, and color intensity. The methodology included experimental measurements of velocity and color intensity using optical methods and image processing.
2:Sample Selection and Data Sources:
Human chorionic gonadotropin (hCG) was used as the analyte, with concentrations of 0, 5, 10, 50, and 250 IU/L. Each test was repeated 10 times for statistical significance.
3:List of Experimental Equipment and Materials:
Equipment included a USB digital microscope (Coolingtech), micropipette (Dragon Lan), Biodot XYZ 3060 Dispensing Platform, manual metal paper cutter, and various pads and membranes from Advance Microdevices PVT LTD. Materials included antibodies, gold nanoparticles, and buffers.
4:Experimental Procedures and Operational Workflow:
Strips were prepared with different widths and test line positions. Sample analyte was applied, and flow velocity was measured using time-lapse imaging. Color intensity was quantified after 10 minutes using RGB processing of images captured by the microscope.
5:Data Analysis Methods:
Data were analyzed using MATLAB for image processing, with statistical significance cutoff of 0.05. Velocity was determined from displacement of fluid front over time.
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USB digital microscope
Coolingtech
Quantifying color intensity by capturing images of test lines and processing RGB values.
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Micropipette
Micopette plus
Dragon Lan
Handling sample liquid during experiments.
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Dispensing Platform
XYZ 3060
Biodot
Printing test line and control line antibodies on nitrocellulose membrane.
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Paper cutter
Ningbo Hollan
Cutting strips to specific trapezoidal profiles.
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Sample pads
GFB-R7L
Advance Microdevices PVT LTD
Part of the lateral flow assay strip for sample application.
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Absorbent pads
AP110
Advance Microdevices PVT LTD
Absorbing sample fluid in the assay strip.
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Conjugate release matrix
PT-R7
Advance Microdevices PVT LTD
Releasing conjugate into the flow in the assay strip.
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Backing cards
L Type
Advance Microdevices PVT LTD
Supporting the porous pads in the assay strip.
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Nitrocellulose test membranes
CNPCSS-12
Advance Microdevices PVT LTD
Serving as the detection pad in the assay strip.
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