RAISING EFFECTIVE POLYCLONAL ANTIBODIES AGAINST PURIFIED PEPPER LEAF CURL VIRUS PARTICLES
Abstract
Sensitive diagnostic tools are urgently needed for rapid and specific virus detection. Pepper Leaf Curl Virus (PeLCV) particles were purified from the injected pepper plants after symptoms manifestation. PeLCV particles were characterized by injection into healthy pepper plants which exhibited typical infection symptoms including curling and downward cup shape of the leaves in 45-50 days post infection. Maintaining PelCV in pepper plants were characterized by PCR using whitefly transmitting geminivirus specific primers which revealed positive signals with DNA extracted from infected pepper plants as a 659 bp DNA fragment. PCR isolated DNA fragment was subjected to sequencing and the determined nucleotide sequences were blasted in the Gene bank databases. Alignments confirmed that the isolated fragment belong to the C1 gene of geminiviruses (core region). Polyclonal antibodies were raised against the purified virus particles and evaluated by ELISA as a trapped and intermediate antibodies as well as western blotting analysis.References
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