The invention concerns the Efa DNA polymerase derived from bacteriophage vB_EfaS-271, capable of fast and efficient isothermal DNA amplification across a broad temperature range from -10°C to 50°C. The enzyme works effectively at room temperature, enabling DNA amplification without specialised laboratory equipment.
The Efa polymerase features 5’-3’ polymerase activity and 3’-5’ proofreading activity, as well as a unique amino-acid sequence (~20 percent identity to other polymerases). In tests, it generated amplification products (~40 bp) within approximately 5 minutes and remained active even below 0°C.
Its production process is simple and energy-efficient, requiring no cooled purification steps. The invention also includes the gene sequence, the pET28-Efa expression vector and an optimised purification procedure. The technology is applicable in diagnostics, pathogen detection, genetic analysis and field-based molecular work.
Application:
- Molecular diagnostics performed outside laboratory settings without a thermocycler,
- Pathogen detection assays based on low-temperature DNA amplification,
- Rapid genetic analyses, genotyping and mutation identification.
