The development of a competitive PCR–ELISA for the detection of equine herpesvirus-1

Daly, Paul; Doyle, Seán

doi:10.1016/s0166-0934(02)00252-5

Cited by 14 publications

(3 citation statements)

References 17 publications

(28 reference statements)

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“…4,8 Recently a nested PCR-enzyme-linked immunosorbent assay (ELISA) system has also been described. 6 All of these techniques suffer from some disadvantages: virus isolation techniques are labor intensive and time consuming; immunohistochemistry and in situ hybridization have limited sensitivity; and conventional PCR and PCR-ELISA are not quantitative.…”

Section: Introductionmentioning

confidence: 99%

Detection and Quantification of Equine Herpesvirus-1 Viremia and Nasal Shedding by Real-Time Polymerase Chain Reaction

et al. 2006

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Equine herpesvirus-1 (EHV-1) infection is common in young horses throughout the world, resulting in respiratory disease, epidemic abortion, sporadic myelitis, or latent infections. To improve on conventional diagnostic tests for EHV-1, a real-time polymerase chain reaction (PCR) technique was developed, using primers and probes specific for the EHV-1 gB gene. Amplification efficiencies of 100% +/- 5% were obtained for DNA isolated from a plasmid, infected peripheral blood mononuclear cells (PBMCs), and nasal secretions from infected ponies. The dynamic range of the assay was 8 log10 dilutions, and the lower limit of detection was 6 DNA copies. Fifteen ponies, seronegative for EHV-1, were experimentally infected with EHV-1, and nasal samples were used to quantify shedding of virus by both virus isolation and real-time PCR analysis. Virus isolation identified nasal shedding of EHV-1 in 12/15 ponies on a total of 25 days; real-time PCR detected viral shedding in 15/15 ponies on 75 days. Viremia was quantified using PBMC DNA, subsequent to challenge infection in 3 additional ponies. Viremia was identified in 1/3 ponies on a single day by virus isolation; real-time PCR detected viremia in 3/3 ponies on 17 days. When real-time PCR was used to analyze PBMC DNA from 11 latently infected ponies (documented by nested PCR), EHV-1 was not detected. We conclude that real-time PCR is a sensitive and quantitative test for EHV-1 nasal shedding and viremia and provides a valuable tool for EHV-1 surveillance, diagnosis of clinical disease, and investigation of vaccine efficacy.

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Section: Introductionmentioning

confidence: 99%

Detection and Quantification of Equine Herpesvirus-1 Viremia and Nasal Shedding by Real-Time Polymerase Chain Reaction

et al. 2006

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“…(The sample should be refrigerated and time minimised before arriving at the laboratory). Tests undertaken may include: z z Quantitative Polymerase Chain Reaction (qPCR) for detecting viral DNA is sensitive and allows estimation of the amount of virus ('viral load') in the sample (Daly and Doyle, 2003;Hussey et al, 2006;). Turn-around is short, with results generally being reported on the day of receipt.…”

Section: Diagnosis Of Ehv Infectionmentioning

confidence: 99%

Equine herpesviruses: a roundtable discussion

Ivens¹,

Rendle²,

Kydd³

et al. 2019

UK-Vet Equine

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Foreword There are nine different equid herpesviruses (EHVs). Five types (EHV-1 to EHV-5) infect the domestic horse, while EHV-6 to EHV-9 are associated with infections in wild equids including asses and zebra. This review focuses on the commonest and most important clinical pathogens, the alphaherpesviruses EHV-1 and 4. These are respiratory pathogens and are also responsible for abortion and neurological disease. Several aspects of the biology of these viruses makes their control challenging. In particular, latent infection and reactivation of infection under stress, with subsequent virus shedding, makes elimination of these viruses impossible. Biosecurity measures are important both for minimising the risk of an outbreak and for controlling any outbreak when it occurs. Recognition of the disease and confirmatory diagnosis are also important in order for appropriate biosecurity measures to be instigated. Vaccination in key demographic groups is also important to reduce severe clinical disease. Unfortunately many horse owners are unaware of EHV or the importance of biosecurity measures and vaccination for control.

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“…''Abortion storms'' were described as early as 1931 as Rift Valley fever swept through herds of cattle and sheep. More recent abortion storms have been attributed to equine herpesvirus type 1 (especially Army 183 F-fetal strain); Coxiella burnetii; Thogoto virus (transmitted by Ixodid ticks); Neospora caninum; and one strain of equine arteritis virus [37][38][39][40][41][42][43][44]. It is important to be aware that abortion storms may be an early sign of a zootic or a bioterrorist attack with a hemorrhagic fever virus or other agents.…”

Section: Phlebovirus Genusmentioning

confidence: 99%

Viral Hemorrhagic Fevers: Current Status of Endemic Disease and Strategies for Control

Cleri

Ricketti

Porwancher

et al. 2006

Infectious Disease Clinics of North America

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The development of a competitive PCR–ELISA for the detection of equine herpesvirus-1

Cited by 14 publications

References 17 publications

Detection and Quantification of Equine Herpesvirus-1 Viremia and Nasal Shedding by Real-Time Polymerase Chain Reaction

Detection and Quantification of Equine Herpesvirus-1 Viremia and Nasal Shedding by Real-Time Polymerase Chain Reaction

Equine herpesviruses: a roundtable discussion

Viral Hemorrhagic Fevers: Current Status of Endemic Disease and Strategies for Control

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