الفهرس 
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Abstract
1. 57 pooled blood samples were collected from clinically affected animals, suggestive of BTV infection, were collected using both plain and EDTA-coated vacutainers.
2. The plain vacutainers (serum) samples were subjected to testing by means of IDEXX Bluetongue antibody competition test.
3. EDTA-coated (whole blood) vacutainers of positive IDEXX samples (Baladi sheep and goats) were used to isolate the buffy coat.
4. The isolated buffy coat was inoculated in 10-day-old SPF-ECE via yolk sac route, for primary isolation, for 9 passages showing hemorrhagic cherry red chicken embryo at the last passage.
5. BTV concentrate was prepared from SPF-ECE homogenate and filtrate using PEG-6000 with high speed centrifugation.
6. The prepared BTV concentrate was examined, for identification, using TEM and showed smaller viral size, but shape typical of BTV.
7.BTV concentrate was employed in hyperimmune serum production in rabbits.
8. Tissue filtrate from SPF-ECE was used for secondary isolation on BHK-21 cell line for 6 passages with appearance of CPE.
9. Immunoelectron transmission microscopy was employed, using control positive BTV anti-serum showing both immunoaggregation and immunolocalization of the antibodies indicating the presence of the virus.
10. TCID50 followed by SNT and direct antigen solid-phase sandwich ELISA were utilized as a means for BTV antigen confirmation and were positive.
11. Viral RNA was extracted, from various biological specimens (blood, serum, SPF-ECE homogenate, BTV concentrate and tissue culture homogenate), using GeneJet Viral DNA and RNA Purification Kit (Thermo Scientific).
12. The extracted RNA’s purity and concentration were assessed using Jenway™ Genova Nano (micro-volume spectrophotometer) and it was considered to be within acceptable limits.
13. Primers targeting highly conserved genes (VP7, VP3, NS1 and NS3) were utilized numerously in RT-PCR experiments for amplification of specific bands, but none was positive.
14. Pooled blood, sera, virus concentrate, tissue and cell culture homogenate were sent to Pirbright Institute (Non-Vesicular Diseases Reference Laboratory – United Kingdom) on FTA cards in another additional trial, using RT-PCR, to explicate the molecular identity of the virus and the analysis result was negative.
15. SPF-ECE homogenate from intrayolk (IY) inoculation was used to perform additional three passages on 13-year-old SPF-ECE using CAM route with appearance of pock lesion.
16. The pock lesion was taken, processed and visualized using TEM revealing the virus existence in tissue.
17. The pooled virus concentrate was sent to Children’s Cancer Hospital – Basic Research Department – Proteomics and Metabolomics Research Program for untargeted shotgun proteomic analysis and the result confirmed the molecular identity of the virus to be BTV.
18. The results prove the presence of a chimeric virus.