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Coupling Sensitive Nucleic Acid Amplification with Commercial Pregnancy Test Strips
Yan Du1, Arti Pothukuchy1, Jimmy D Gollihar1
1Institute for Cellular and Molecular Biology, Center for Systems and Synthetic Biology, Department of Chemistry, The University of Texas at Austin, Austin, TX, 78712, USA.
Angewandte Chemie (International Ed. in English)
|December 20, 2016
Summary
This study presents a novel point-of-care diagnostic method using isothermal amplification and pregnancy test strips. The assay enables rapid, low-cost detection of nucleic acid biomarkers, including Ebola virus and melanoma-associated SNPs.
Area of Science:
- Biotechnology
- Molecular Diagnostics
- Point-of-Care Testing
Background:
- Current point-of-care (POC) nucleic acid biomarker detection faces challenges in technical complexity, cost, and time.
- Existing methods often require sophisticated equipment and trained personnel, limiting widespread POC application.
Purpose of the Study:
- To develop a simplified, cost-effective, and rapid POC diagnostic assay for nucleic acid biomarkers.
- To integrate loop-mediated isothermal amplification (LAMP) with toehold-mediated strand-exchange and pregnancy test strips for signal transduction.
Main Methods:
- Combined loop-mediated isothermal amplification (LAMP) with programmable toehold-mediated strand-exchange signal transduction.
- Utilized an engineered hCG-SNAP fusion reporter protein for signal amplification and transduction.
- Adapted the system to standard, low-cost, commercially available pregnancy test strips for readout.
Main Results:
- Successfully detected as few as 20 copies of Ebola virus templates in human serum and saliva.
- Demonstrated adaptability for distinguishing a melanoma-associated SNP allele (BRAF V600E) from wild-type sequences.
- Achieved reliable signal transduction from LAMP amplification to a visible readout on pregnancy test strips.
Conclusions:
- The developed method offers a versatile and generalizable platform for POC nucleic acid biomarker detection.
- This approach overcomes limitations of current POC diagnostics, enabling rapid and cost-effective testing for various pathogens and genetic markers.
- The technology holds significant potential for broad applications in infectious disease diagnostics and personalized medicine.
