Development and Evaluation of a Real-time Polymerase Chain Reaction Assay for Equine Encephalosis Virus
نویسنده
چکیده
........................................................................................................................................... IX 1. GENERAL INTRODUCTION .......................................................................................................... 1 2. LITERATURE REVIEW ................................................................................................................... 3 2.1. Classification and characterization of EEV .............................................................................. 3 2.2. Structural characteristics ......................................................................................................... 5 2.3. Viral genome composition ....................................................................................................... 5 2.4. Structural proteins ................................................................................................................... 6 2.5. Non-structural proteins ............................................................................................................ 8 2.6. Epidemiology ......................................................................................................................... 10 2.7. Clinical signs and pathogenesis ............................................................................................ 11 2.8. Diagnosis ............................................................................................................................... 13 3. MATERIALS AND METHODS ...................................................................................................... 14 3.1. Primer design ........................................................................................................................ 14 3.2. EEV Isolates .......................................................................................................................... 14 3.3. RNA Extractions .................................................................................................................... 15 3.4. Reverse transcription polymerase chain reaction (RT-PCR) ................................................ 15 3.5. Sequencing ............................................................................................................................ 16 3.6. Sequence analyses ............................................................................................................... 16 3.7. Assay characterization .......................................................................................................... 17 4. RESULTS ...................................................................................................................................... 18 4.1. Primer design ........................................................................................................................ 18 4.2. Reverse transcription polymerase chain reaction (RT-PCR) ................................................ 18 4.3. Sequencing ............................................................................................................................ 18 4.4. Primers and TaqMan® MGB hydrolysis probe ................................................................... 19 4.5. Real time reverse transcription polymerase chain reaction (rtRT-PCR) ............................... 23 5. DISCUSSION AND CONCLUSIONS ............................................................................................ 25 6. REFERENCES .............................................................................................................................. 27 7. APPENDIX .................................................................................................................................... 31
منابع مشابه
Development of a real time polymerase chain reaction assay for equine encephalosis virus.
Equine encephalosis virus (EEV) is the cause of equine encephalosis. The disease is similar to mild forms of African horse sickness (AHS) and the two diseases are easily confused. Laboratory identification and serotyping of EEV is based on viral isolation in BHK-21 cells and a viral plaque inhibition neutralisation test. These procedures are time-consuming and therefore a more rapid diagnostic ...
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