Development of isothermal Recombinase polymerase amplification (RPA) process for detection of Leptospira spp. causing diseases

Abstract

Leptospirosis is one of common infectious diseases in the world with the pathogen as Leptospira spp. belonging to Leptospira. In Vietnam, leptospirosis is considered a endemic disease, and there is a high risk of outbreaks if not diagnosed and treated promptly. Application of realtime PCR technique is promoting technique in the detection of Leptospira spp. at the early stages of the disease. However, this technique is too complex and demanding in the operation of the thermal cycler, so it is increasingly replaced by isothermal amplification methods. In this study, the team successfully developed an isothermal amplification process using Reacombinase polymerase amplification (RPA) to accurately detect Leptospira spp. causing the disease with the detection time of only 30 minutes. The RPA process developed by the team not only has a limit of detection (LOD) equivalent to the commercially available kit (100 copies/reaction) but also achieves similar sensitivity and specificity (100%).