Equine herpesvirus 1 (EHV-1) nucleotide polymorphism determination using formalin fixed tissues in EHV-1 induced abortions and myelopathies with real-time PCR and pyrosequencing |
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Authors: | Deepanker Tewari Fabio Del Piero Stephen Cieply Willard Feria Helen Acland |
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Institution: | 1. Pennsylvania Veterinary Laboratory, Pennsylvania Department of Agriculture, 2305 North Cameron Street, Harrisburg, PA, United States;2. Department of Pathobiology, University of Pennsylvania, 382 West Street Road, Kennett Square, PA, United States |
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Abstract: | Equine herpesvirus-1 (EHV-1) strains with a single point mutation at the 2254 nucleotide position with a G2254 constitution within the DNA polymerase gene are associated strongly with equine myeloencephalopathies. Infections with non-neuropathogenic EHV-1 strains without the G2254 nucleotide but with an A2254 nucleotide are associated less frequently with equine neurologic disease. A retrospective study utilizing DNA extracted from formalin fixed paraffin embedded tissues was conducted with real time PCR and pyrosequencing, to determine the infecting EHV-1 strains. Infection with EHV-1 A2254 and or G2254 strain was detected with real time PCR, and was confirmed with a rapid pyrosequencing technique. Pyrosequencing was useful in at least 2 cases where real time PCR was equivocal in determining the infecting EHV-1 strain type. The strain with G2254 mutation was detected in 9.4% of 21 studied abortion cases, and in 86.6% of 15 neurologic cases. |
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Keywords: | Equine herpesvirus Pyrosequencing Neuropathogenic Non-neuropathogenic Nucleotide polymorphism A/G2254 |
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