DEVELOPMENT OF A REVERSE TRANSCRIPTION-LOOP MEDIATED ISOTHERMAL AMPLIFICATION ASSAY FOR RAPID DETECTION OF INFECTIOUS BURSAL DISEASE VIRUS IN CHICKENP. RAJA, T.M.A. SENTHILKUMAR, M. PARTHIBAN, A. THANGAVELU AND A. MANGALA GOWRIAbstract Infectious bursal disease (IBD) is an immunosuppressive disease caused by infectious bursal disease virus (IBDV) that affects chickens, results in significant economic losses to the poultry industry worldwide. With the objective of developing a simple and rapid method to detect IBDV, a reverse transcription-loop mediated isothermal amplification assay (RT-LAMP) was developed and evaluated in this study. This method targets the well-conserved RNA dependent RNA polymerase (VP1) gene of IBDV and has the potential to detect diverse IBDV isolates. RT-LAMP amplified products were successfully detected by naked eye using SYBR Green I and when subjected to agarose gel electrophoresis revealed ladder-like pattern. The sensitivity of the RT-LAMP was compared with other genome amplification methods such as RT-PCR and real time RT-PCR methods. It was observed that RT-LAMP was more sensitive than RT-PCR but as sensitive as real time PCR. Furthermore, this method exhibited excellent specificity without any cross reaction to other common avian pathogens. This method also exhibited superior practical applicability when tested on tissue samples suspected for IBDV. Overall, the IBDV VP1-gene based RT-LAMP assay developed in this study is a rapid, simple and sensitive assay suitable for less-equipped laboratories and can be utilized in field conditions as a screening test for rapid detection of IBDV. |