A Novel PCR Method for Detecting ACE Gene Insertion/Deletion Polymorphisms and its Clinical Application.

Area of Science:

• Biotechnology
• Molecular Diagnostics
• Genetics

Background:

• Angiotensin-converting enzyme (ACE) is crucial for blood pressure regulation and cardiovascular homeostasis.
• ACE's diverse functions suggest its involvement in various diseases.

Purpose of the Study:

• To develop a novel, rapid visual detection method for ACE I/D polymorphisms.
• To enable direct genotyping from clinical samples without prior DNA extraction.

Main Methods:

• Integration of direct Polymerase Chain Reaction (PCR) with gold magnetic nanoparticles (GMNPs)-based lateral flow assay (LFA) biosensor.
• Elimination of the DNA extraction step for streamlined sample processing.
• Validation using sequencing on 300 peripheral blood samples and a preliminary clinical study on 633 samples.

Main Results:

• The developed direct PCR-LFA system provides ACE I/D genotyping in approximately 80 minutes.
• The assay has a detection limit of 0.75 ng, with results available in 5 minutes.
• High accuracy and repeatability were confirmed through comparison with sequencing.

Conclusions:

• A significant association between ACE genotypes and patient age was observed in the Coronary Heart Disease (CHD) group.
• This direct PCR-LFA method shows potential for clinical application in diagnosing diseases linked to large fragment gene insertions/deletions.