研究目的
To develop a point-of-care diagnostic test for quantifying levels of folate in human serum to improve understanding of folate status in women of reproductive age and reduce the risk of neural tube defects.
研究成果
The developed point-of-care diagnostic test accurately quantifies folate in human serum, offering a rapid, inexpensive solution for assessing folate status in resource-limited settings. This technology has the potential to improve global maternal and fetal healthcare through enhanced understanding of folate status in women of reproductive age.
研究不足
The assay requires serum to be previously separated from whole blood, and future work is needed to test the assay with unaltered serum samples containing endogenous folate and other serum components.
1:Experimental Design and Method Selection:
The study employs a lateral flow assay combined with a portable imaging device for folate quantification.
2:Sample Selection and Data Sources:
Folate-free human serum was prepared and spiked with known concentrations of folic acid and 5-MTHF for testing.
3:List of Experimental Equipment and Materials:
Includes plastic-backed nitrocellulose membranes, glass fiber conjugate pads, cellulose fiber sample pads, fluorescein isothiocyanate (FITC) conjugation kits, folic acid-BSA conjugates, anti-FITC antibodies, bovine folate binding protein (FBP), and a portable imaging device.
4:Experimental Procedures and Operational Workflow:
The lateral flow assay assembly involves dispensing test and control lines onto nitrocellulose membranes, preparing conjugate pads, and assembling the lateral flow strips. The assay procedure includes applying serum samples and running buffer to the sample pad, followed by imaging the strip with the portable device.
5:Data Analysis Methods:
The ratio of test line to control line fluorescence intensity is calculated to determine folate concentration.
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Plastic-backed nitrocellulose membranes
HF180MC100
EMD Millipore
Used as the base material for the lateral flow assay.
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Glass fiber conjugate pads
GFDX103000
EMD Millipore
Used in the assembly of the lateral flow assay.
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Cellulose fiber sample pads
CFSP203000
EMD Millipore
Used for sample application in the lateral flow assay.
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Fluorescein isothiocyanate (FITC) Lightning Link Conjugation Kits
Innova Biosciences
Used for fluorescent labeling in the assay.
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Folic acid-BSA conjugates
Fitzgerald Industries
Used for binding to FITC in the assay.
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Anti-FITC antibodies
Abcam Inc.
Used for the control line in the assay.
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Bovine folate binding protein (FBP)
Scripps Laboratories
Used for the test line in the assay.
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Portable imaging device
Used to image and analyze the lateral flow assay strips.
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