Abstract
Foot and mouth disease (FMD) - list-A disease - is the major constraint in international trade of livestock and dairy products. FMD virus exist in the nature as seven distinct serotypes i.e A, O, C, Asia-I, SAT 1-3. There is considerable heterogeneity within serotypes and there is no cross immunity between serotypes, Hence, diagnosis of this disease up to strain level is always a problem. The ultimate solution for this problem is Reverse-Transcription Polymerase Chain Reaction (RT-PCR). In the present study, 156 samples were collected from FMD suspected animals from seven different districts (Jhang, Faisalabad, Lahore, Hafizabad, Okara, Gujranwala and Arifwala) of the Punjab province of Pakistan. For this purpose, vesicles from mouth and hooves and tongue tissues of dead animals were collected and subjected to RT-PCR analysis. RNA was extracted using TRIzolTM reagent and subsequently subjected to cDNA synthesis. The cDNA was amplified through PCR using FMD virus (FMDV) specific universal primers P1 and P2, which correspond to highly conserved region of the viral genome i.e VP1, to achieve the required band size of 216 bp. Thirty-six (23%) samples were found positive following RT-PCR analysis. The optimization of this molecular biological technique (RT-PCR) will be of value for the accurate diagnosis and differential diagnosis of FMD.
