Nucleic Acid Amplification using Recombinase Polymerase: Enzymatic Approach

Abstract

Rapid detection of infections is crucial for the prevention of infectious disease outbreaks, development of anti-microbial drugs and biodefense. When considering a diagnostic test the most important considerations are rapidity, ease of use, portability, specificity and sensitivity. Recently developed isothermal recombinase polymerase amplification (RPA) technology has been shown in many publications to be the most sensitive and effective for determining infections, require no sophisticated and expensive equipment, and is suitable for point-of-care field applications. Here we have described our viewpoints with regard to RPA technology’s suitability and usage in resource poor settings, its advantages and limitations. We have also developed a lateral flow assay to detect all serotypes of dengue virus following RPA procedure, demonstrating its suitability for field applications. We believe our suggestions may help in improving RPA procedures and may also help in transitioning to clinical applications.