Production of specific antisera against Beet mosaic virus and Beet necrotic yellow vein virus
Production of specific antisera against Beet mosaic virus and Beet necrotic yellow vein virus
Allam A. Megahed1; Hoda M.A. Waziri2; Khaled A. El-Dougdoug3; Badawi A. Othman3; Sirag M. Lashin1; Mohamed D. Hassanin1 and Mahmoud A. Ibrahim4
ABSTRACT
The ultraviolet spectrum of purified Beet mosaic virus (BtMV) particles obtained by density gradient centrifugation revealed that, the concentration of virus preparation estimated using spectrophotometric measurements at 260 nm was 3.004 mg/ml. The min, max nm, Amax./Amin., A260/A280 and A280/A260 ratios were 244, 260, 1.276, 1.512 and 0.662, respectively for BtMV. The corresponding figures of the purified virus preparation of Beet necrotic yellow vein virus (BNYVV) were 2.196 mg/ml, and the min, max nm, Amax./Amin., A260/A280 and A280/A260 were 248, 264, 1.606, 1.237, and 0.785, respectively. The polyclonal antibodies raised against BtMV and BNYVV by 5 injection doses each, using different injection types obtained from rabbit bleedings 10 days after the last injection had the titer of 1:2048 and 1:1024, respectively by indirect-enzyem linked immunosorbent assay (I-ELISA) technique. The produced antisera were evaluated and compared with foreign antisera using tissue printing immunoassay (TPIA) and dot blot immunoassay (DBIA). The produced antisera were found more efficiency in development the purplish blue color than the positive reaction when the foreign antisera applied, therefore it should be use produced antisera from Egyptian isolates specific for detection of sugar beet viral infections as a good replacement of foreign ones.
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