Development And Assessment Of Loop-mediated Isothermal Amplification For Detection Of Foot-and-mouth Disease Virus

Food-and-Mouth disease (FMD) is a highly fatal and contagious disease caused by Food-and-Mouth disease virus (FMDV). It mainly infects cloven-hoofed animals and sometimes can be found in human and other animals. As a single-stranded RNA virus, FMDV has highly variability and its antigenity changes much frequently, so it is much difficult to identify and control the disease.At present, there are many methods to detect Food-Mouth disease virus (FMDV) from blood, serum or tissues, such as virus isolation and characterization, immunological tests, serological test, molecular biological examination, and so on. These detecting methods have supplied powerful technology for the detection of FMDV, but some new molecular technologies, such as loop-mediated isothermal amplification (LAMP), are developed in the recent years and thus supply new skills for the quick and specific examination of FMD pathogens.Aimed to develop a quick diagnostic method of FMDV, The 3D genes of foot-and-mouth virus type Asiaâ… were downloaded from GeneBank and analyzed using MegAlign tools of software DNAStar. The outer, inner and loop primers were then designed in their conservative region using software PrimerExplorer V3. The LAMP method of detecting foot-and-mouth virus was set up and optimized to be sensitive and specific for the examination of FMDV from tissues. The fragment of foot-and-mouth virus sequence was also amplified and cloned into promega T easy vector to serve as the positive standard and optimized its LAMP reaction condition and reaction system in this experiment. The results showed that the LAMP method developed in this experiment was with high specificity and sensitivity, and the deveolped LAMP method was as highly sensative as REAL-time PCR and can meet the demands of clinical detection of FMDV contamination.