Highly sensitive and rapid detection of porcine circovirus 2 by avidin-biotin complex based lateral flow assay coupled to isothermal amplification

Abstract

In this study, a new platform for the detection of porcine circovirus 2 was developed by avidin-biotin complex based lateral flow assay (LAMP-LFA). Improved detection sensitivity was attained by using loop mediated isothermal amplification (LAMP) with a low limit of detection (LOD), so the platform can be used to detect even early or asymptomatic stages of infection. LFA, which requires no specialized equipment, facilitates the use of point-of-care (POC) tests. Therefore, by applying LFA, the result can be confirmed accurately with the naked eye. Moreover, this platform has a unique structure using a single-tag detection system. The avidin-biotin interaction is the strongest interaction between proteins and has a higher K-d value than antigen-antibody interactions. Thus, the results are stable and can be clearly confirmed. The high sensitivity of LAMP-LFA enables all steps to be completed in 30 min. As a result, it could be applied to different targets, such as other pathogens. Future POC diagnostic studies are expected to be of great practical benefit.