Visual Detection of ACE I/D Polymorphism Using T-ARMS-PCR Combined with a Lateral Flow Assay and Its Clinical...

Area of Science:

• Molecular Biology
• Genetics
• Biotechnology

Background:

• Insertion/deletion (indel) variations are key disease markers.
• Current indel genotyping methods are complex, time-consuming, and costly.
• There is a need for rapid, point-of-care diagnostic tools.

Purpose of the Study:

• To develop a fast and accurate biosensor for visual indel genotyping.
• To enable point-of-care testing for genetic variations.
• To simplify the process of genotyping specific large indel variants.

Main Methods:

• Developed a biosensor combining tetra-primer amplification refractory mutation system polymerase chain reaction (T-ARMS-PCR) and GoldMag lateral flow assay (LFA).
• Utilized four primers in a single PCR reaction for distinguishing ACE I/D polymorphism.
• Visual genotyping results obtained within 5 minutes via LFA strip color changes.

Main Results:

• The T-ARMS-PCR-LFA system accurately genotyped ACE I/D polymorphism in 50 human DNA samples with 100% accuracy.
• Successful genotyping was achieved with a low DNA input (25 ng).
• The method eliminated complex operation steps and data analysis compared to conventional methods like agarose gel electrophoresis.

Conclusions:

• The T-ARMS-PCR-LFA biosensor provides a rapid, accurate, and visually interpretable method for indel genotyping.
• This technology is highly applicable for clinical practices, enabling faster decision-making in disease diagnosis.
• The biosensor facilitates personalized medicine by simplifying genetic variant analysis.