Purification and antiserum production of banana bract mosaic poty virus in the Philippines
2004
Villamor, D.E.V. | Herradura, L.E. | Bajet, N.B. (Lapanday Agricultural Development Corp., Mandug, Davao City (Philippines). Research Dept.)
Banana bract mosaic potyvirus (BBrMV) was extracted and purified from field collected, BBrMV-ELISA positive banana leaves using virus antiserum from Australia by following the procedure of Thomas et al (1997). Rate zonal cesium chloride gradient centrifugation of the partially purified preparation showed one distinct light scattering band and this band was collected. Assays of its aliquots showed that it has a typical nucleoprotein profile at UV 220-320 nm, reacted with the same antiserum in ELISA and showed major proteins of about 31 and 39 kDa in SDS-PAGE [sodium dodecyl sulfate-polyacrylamide gel electrophoresis]. These proteins showed strong signals with the same antiserum in Western blot. Low virus yield was obtained but the concentration was enough to produce virus antiserum in rabbits. After immunizing two rabbits with the purified virus preparations, only one produced antiserum (R2 BBrMV-As) with relatively sufficient titer and of comparable reaction with the antiserum from Australia in both ELISA and western blot. The successful purification demonstrates local capability of obtaining relatively pure samples for more fundamental studies on the virus particle. The availability of this R2 BBrMV-As complements and expands existing antibody-based assays for the detection of this virus in banana and other host plants, thus, enhancing diagnosis of virus diseases of Musa spp. in the country
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