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Development of a reverse transcriptasepolymerase chain reaction assay to detect feline herpesvirus-1 latency-associated transcripts in the trigeminal ganglia and corneas of cats that did not have clinical signs of ocular disease

Wendy M. TownsendDepartment of Veterinary Clinical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907-2026.
Present address is the Department of Small Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University, East Lansing, MI 48824-1314.

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Jean StilesDepartment of Veterinary Clinical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907-2026.

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Lynn Guptill-YoranDepartment of Veterinary Clinical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907-2026.

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Sheryl G. KrohneDepartment of Veterinary Clinical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907-2026.

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Abstract

Objective—To develop a reverse transcriptase-polymerase chain reaction (RT-PCR) assay to detect feline herpesvirus-1 (FHV-1) latency-associated transcripts (LATs) in the corneas and trigeminal ganglia of cats that did not have clinical signs of ocular disease.

Sample Population—Corneas and trigeminal ganglia obtained from 21 cats necropsied at the Indiana Animal Disease Diagnostic Laboratory and 25 cats euthanatized at a humane shelter; none of the cats had a recent history of respiratory tract or ocular disease, and all had normal results for ophthalmic examinations.

Procedure—Both corneas and both trigeminal ganglia were harvested from each cat. An initial PCR assay detected FHV-1 DNA in the corneas and trigeminal ganglia. The RNA was then isolated from samples positive for FHV-1 DNA, and an RT-PCR assay was used to detect LATs.

Results—FHV-1 DNA was detected in 45 of 92 (48.9%) corneas and 38 of 92 (41.3%) trigeminal ganglia. In many samples, the RNA had degraded and RTPCR assay was not possible. Of the samples subjected to RT-PCR assay, none of the 39 corneas but 4 of 16 trigeminal ganglia had positive results when tested for LATs.

Conclusions and Clinical Relevance—Analysis of the results indicated that a high percentage of cats that did not have clinical signs of ocular disease had detectable FHV-1 DNA in their corneas and trigeminal ganglia. This study documents that the RT-PCR assay can successfully identify LATs and may serve as a tool to better understand the biologic characteristics of FHV-1 and its relationship to clinical disease. ( Am J Vet Res 2004;65:314–319)

Abstract

Objective—To develop a reverse transcriptase-polymerase chain reaction (RT-PCR) assay to detect feline herpesvirus-1 (FHV-1) latency-associated transcripts (LATs) in the corneas and trigeminal ganglia of cats that did not have clinical signs of ocular disease.

Sample Population—Corneas and trigeminal ganglia obtained from 21 cats necropsied at the Indiana Animal Disease Diagnostic Laboratory and 25 cats euthanatized at a humane shelter; none of the cats had a recent history of respiratory tract or ocular disease, and all had normal results for ophthalmic examinations.

Procedure—Both corneas and both trigeminal ganglia were harvested from each cat. An initial PCR assay detected FHV-1 DNA in the corneas and trigeminal ganglia. The RNA was then isolated from samples positive for FHV-1 DNA, and an RT-PCR assay was used to detect LATs.

Results—FHV-1 DNA was detected in 45 of 92 (48.9%) corneas and 38 of 92 (41.3%) trigeminal ganglia. In many samples, the RNA had degraded and RTPCR assay was not possible. Of the samples subjected to RT-PCR assay, none of the 39 corneas but 4 of 16 trigeminal ganglia had positive results when tested for LATs.

Conclusions and Clinical Relevance—Analysis of the results indicated that a high percentage of cats that did not have clinical signs of ocular disease had detectable FHV-1 DNA in their corneas and trigeminal ganglia. This study documents that the RT-PCR assay can successfully identify LATs and may serve as a tool to better understand the biologic characteristics of FHV-1 and its relationship to clinical disease. ( Am J Vet Res 2004;65:314–319)