PJB-2005-51
COMPARISON OF PURIFICATION METHODS EMPLOYED FOR PAKISTANI ISOLATE OF BANANA BUNCHY TOP VIRUS
TAHIRA YASMIN, HUSSAIN SHAH AND SAIF KHALID
Abstract
Banana Bunchy Top Virus (BBTV) particles were purified employing various purification methods. Substantial amount of virus particles were obtained using tangential flow filters of millipore sizes of 10 and 0.16µm and 100KDa/100mwco sequentially. The method was most successful and gave high virus yield (75µg/ml) devoid of most host macromolecules. Under electron microscopy (EM) isometric virus particles of 18-20 nm in diameter were observed. The purified virus was used to raise polyclonal antiserum against local isolate of BBTV giving an antibody titre of 1:6000 higher than other methods. Later Enzyme-linked immunosorbent assay (ELISA) was also standardized for BBTV indexing.
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