title: 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.
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.
authors.affiliation: Pennsylvania Veterinary Laboratory, Pennsylvania Department of Agriculture, 2305 North Cameron Street, Harrisburg, PA, United States. Electronic address: dtewari@pa.gov.
authors.forename: Deepanker
authors.initials: D
authors.lastname: Tewari
authors.forename: Fabio
authors.initials: F
authors.lastname: Del Piero
authors.forename: Stephen
authors.initials: S
authors.lastname: Cieply
authors.forename: Willard
authors.initials: W
authors.lastname: Feria
authors.forename: Helen
authors.initials: H
authors.lastname: Acland
mesh_terms: D000022:Abortion, Spontaneous / Q000821:virology*; D000818:Animals; D006566:Herpesviridae Infections / Q000150:complications / Q000662:veterinary* / Q000821:virology; D004861:Herpesvirus 1, Equid / Q000235:genetics* / Q000302:isolation & purification; D006734:Horse Diseases / Q000821:virology*; D006736:Horses; D057089:Pathology, Molecular / Q000379:methods*; D011110:Polymorphism, Genetic; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012189:Retrospective Studies; D017422:Sequence Analysis, DNA / Q000379:methods; D013118:Spinal Cord Diseases / Q000821:virology*; D016707:Tissue Fixation
keywords: A/G(2254); Equine herpesvirus; Neuropathogenic; Non-neuropathogenic; Nucleotide polymorphism; Pyrosequencing