STANDARDIZATION OF MULTIPLEX REVERSE TRANSCRIPTION-POLYMERASE CHAIN REACTION AND TYPING OF FOOT-AND-MOUTH DISEASE VIRUS PREVALENT IN BANGLADESH
Keywords:Foot-and-mouth disease virus, serotypes, multiplex RT-PCR
Foot-and-mouth disease (FMD) is a devastating viral disease of cattle that causes severe economic losses in terms of loss of production and calf mortality in Bangladesh. Despite of regular vaccination, outbreak of the disease has become a regular event throughout the country every year. Determination of prevailing serotypes of the causal agent foot-and-mouth disease virus (FMDV) is now crucial need for strategic vaccination programme. The present research work was aimed to standardize a multiplex RT-PCR assay typing of foot-and-mouth disease virus serotypes prevalent among cattle population of Bangladesh. Uniplex and multiplex RT-PCRs were successfully developed and standardized using the extracted RNA of reference FMDV (Type A, O and Asia 1) following adjustment of the concentration of the viral RNA of each serotype, volume of reaction mixture and thermal profile. The mPCR was evaluated on 82 field samples (vesicular fluid, tongue epithelium and tissue from inter-digital space) of the years 2007 and 2008. Of the 82 field samples, 56 (68.29%) were found positive for FMDV. The mPCR successfully differentiated single as well as dual serotypes infection. The serotypes A, O and Asia 1 were confirmed in the samples of the year 2007 and only serotype O in samples of the year 2008. Higher detection rate was found in vesicular fluid (100%) followed by tongue epithelium (79.66%). It may be concluded that the MRT-PCR standardized in this study could be used for detection and differentiation of FMDV serotypes using field samples.
Bangl. J. Vet. Med. (2010). 8 (2) : 149-155