Standardization and Comparison of Loop mediated isothermal amplification with conventional PCR on MD positive archived samples

Marek’s disease is a Lymphoprliferative disease of poultry caused by alphaherpesvirus. Despite the ubiquitous use of vaccination to control losses, MD still affects a poultry industry with an estimated annual loss up to US $2 billion. Therefore, it is essential to better the ability of detection of MD virus during the early stages of an outbreak. The aim of the study was to develop a loop-mediated isothermal amplification (LAMP) method for the simple, rapid and inexpensive detection of MDV from the tissues of MD positive birds and to compare with diagnostic efficacy of conventional PCR. The LAMP was established using a set of three pair of primers specific to MEQ gene for detecting serotype 1 MDV. The amplification was able to finish in 60 min under isothermal condition at 65oC by addition of Bst polymerase. The results were visualized directly under daylight or ultraviolet light by addition of SYBR Green I dye. The Assay demonstrate a significant advantage of loop-mediated isothermal amplification in diagnosis of Marek's disease compared to conventional PCR as it can be carried out in field condition where rapid diagnosis is required.