2.50
Hdl Handle:
http://hdl.handle.net/2336/30776
Title:
Polymerase chain reaction for laboratory diagnosis of orf virus infections
Authors:
Torfason, Einar G; Gudnadottir, Sigrun
Citation:
J. Clin. Virol. 2002, 24(1-2):79-84
Issue Date:
1-Feb-2002
Abstract:
BACKGROUND: The orf virus of sheep and goats is one of several zoonotic parapoxviruses. In the ovine/caprine host it causes contagious ecthyma (contagious pustular dermatitis, scabby mouth), but in humans it normally causes solitary or clustered orf lesions, typically on hands, arms or face. In addition to disease in the animals, the virus can be quite a nuisance as an occupational hazard in farmers and butchers. Clinical diagnosis is often possible, but laboratory diagnosis is sometimes necessary. For virus isolation, primary ovine or bovine cells, not routinely present, are needed. Serological methods exist, but electron microscopy is the most commonly used method. OBJECTIVES: To develop a reliable method for the laboratory diagnosis of orf zoonoses, without virus culture and without access to an electron microscope. STUDY DESIGN: A suitable primer pair was designed for orf polymerase chain reaction (PCR), using the Oligo software and sequence information from GenBank. Orf positive controls and specimens were kindly provided by several public health centers. Molluscum contagiosum specimens were provided by a dermatologist. HSV-1, HSV-2 and VZV positive swab specimens came from our routine diagnostic service. Asymptomatic skin specimens were obtained from sheep heads from the abattoir, and swab specimens from the heads of asymptomatic sheep. Selected amplified orf PCR positive specimens were sequenced to ensure the authenticity of the PCR products. Orf positive specimens were sent to another laboratory for electron microscopy. RESULTS AND CONCLUSIONS: A robust PCR was developed, with very small inter-run variation. All specificity demands were met, and the sensitivity seems to be good or excellent. All negative specificity controls from cell cultures and non-orf viruses were negative. Twenty-two (95.7%) of 23 scab or swab specimens with suspected orf etiology were orf PCR positive. Five of eight skin specimens from sheep heads from the abattoir were positive, and all 11 swab specimens from asymptomatic sheep were negative. Electron microscopy demonstrated orf-like particles in orf-PCR positive specimens. This PCR seems to be suitable as a diagnostic test for orf in humans, but asymptomatic virus shedding in sheep or goats may complicate veterinary applications of the assay.
Description:
Neðst á síðunni er hægt að nálgast greinina í heild sinni með því að smella á hlekkinn View/Open
Additional Links:
http://www.sciencedirect.com/science/article/B6VJV-44MFSG6-D/1/622e5de7502817dec6d803b4b6661b01

Full metadata record

DC FieldValue Language
dc.contributor.authorTorfason, Einar G-
dc.contributor.authorGudnadottir, Sigrun-
dc.date.accessioned2008-07-01T16:11:13Z-
dc.date.available2008-07-01T16:11:13Z-
dc.date.issued2002-02-01-
dc.date.submitted2008-07-01-
dc.identifier.citationJ. Clin. Virol. 2002, 24(1-2):79-84en
dc.identifier.issn1386-6532-
dc.identifier.pmid11744431-
dc.identifier.doi10.1016/S1386-6532(01)00232-3-
dc.identifier.urihttp://hdl.handle.net/2336/30776-
dc.descriptionNeðst á síðunni er hægt að nálgast greinina í heild sinni með því að smella á hlekkinn View/Openen
dc.description.abstractBACKGROUND: The orf virus of sheep and goats is one of several zoonotic parapoxviruses. In the ovine/caprine host it causes contagious ecthyma (contagious pustular dermatitis, scabby mouth), but in humans it normally causes solitary or clustered orf lesions, typically on hands, arms or face. In addition to disease in the animals, the virus can be quite a nuisance as an occupational hazard in farmers and butchers. Clinical diagnosis is often possible, but laboratory diagnosis is sometimes necessary. For virus isolation, primary ovine or bovine cells, not routinely present, are needed. Serological methods exist, but electron microscopy is the most commonly used method. OBJECTIVES: To develop a reliable method for the laboratory diagnosis of orf zoonoses, without virus culture and without access to an electron microscope. STUDY DESIGN: A suitable primer pair was designed for orf polymerase chain reaction (PCR), using the Oligo software and sequence information from GenBank. Orf positive controls and specimens were kindly provided by several public health centers. Molluscum contagiosum specimens were provided by a dermatologist. HSV-1, HSV-2 and VZV positive swab specimens came from our routine diagnostic service. Asymptomatic skin specimens were obtained from sheep heads from the abattoir, and swab specimens from the heads of asymptomatic sheep. Selected amplified orf PCR positive specimens were sequenced to ensure the authenticity of the PCR products. Orf positive specimens were sent to another laboratory for electron microscopy. RESULTS AND CONCLUSIONS: A robust PCR was developed, with very small inter-run variation. All specificity demands were met, and the sensitivity seems to be good or excellent. All negative specificity controls from cell cultures and non-orf viruses were negative. Twenty-two (95.7%) of 23 scab or swab specimens with suspected orf etiology were orf PCR positive. Five of eight skin specimens from sheep heads from the abattoir were positive, and all 11 swab specimens from asymptomatic sheep were negative. Electron microscopy demonstrated orf-like particles in orf-PCR positive specimens. This PCR seems to be suitable as a diagnostic test for orf in humans, but asymptomatic virus shedding in sheep or goats may complicate veterinary applications of the assay.en
dc.language.isoenen
dc.publisherElsevier Scienceen
dc.relation.urlhttp://www.sciencedirect.com/science/article/B6VJV-44MFSG6-D/1/622e5de7502817dec6d803b4b6661b01en
dc.subject.meshAnimalsen
dc.subject.meshBase Sequenceen
dc.subject.meshDNA, Viralen
dc.subject.meshEcthyma, Contagiousen
dc.subject.meshHumansen
dc.subject.meshMolecular Sequence Dataen
dc.subject.meshOrf virusen
dc.subject.meshPolymerase Chain Reactionen
dc.subject.meshSensitivity and Specificityen
dc.subject.meshSheepen
dc.subject.meshSkinen
dc.subject.meshZoonosesen
dc.titlePolymerase chain reaction for laboratory diagnosis of orf virus infectionsen
dc.typeArticleen
dc.contributor.departmentDepartment of Medical Virology, Landspítali-University Hospital, PO Box 8733, IS-128 Reykjavik, Iceland. einarg@landspitali.isen
dc.identifier.journalJournal of clinical virology : the official publication of the Pan American Society for Clinical Virologyen

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