Cloning of the 2B Region from Foot-and-Mouth Disease Viral Genome as a Potential External Positive Control for FMD Detection

Abstract

Foot and Mouth Disease (FMD) is a concern in many tropical and developing countries and pose a continuous threat in FMD-free nations such as Japan, Indonesia, and Singapore. A recent method being used to detect the virus is an RNA-based FMD detection kit. However, RNA can be easily degraded accounting to its shorter shelf life. In this study, RNA isolate from an inactivated vaccine, instead of the live FMD virus, was reverse transcribed and cloned to construct a more stable plasmid DNA. This innovation generated a template that can be stored for a long period and be used as a potential external positive control for DNA-based FMD detection kits. This study proves that the recombinant molecule containing 2B region of FMD virus genome can be used as an external positive control for RTPCR-based FMD detection and can be incorporated in test kits. Moreover, the constructed plasmid is stable for use after five months. Thus, there is no need to extract RNA from live FMD virus which makes the routine faster, convenient, and practical.