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Detection and quantitation of gallid herpesvirus 1 in avian samples by 5′ taq nuclease assay utilizing minor groove binder technology

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Corney, B.G., Diallo, I.S., Wright, L.L., de Jong, A.J., Hewitson, G.R., Tolosa, M.X., Rodwell, B.J., Ossedryver, S. M., Pritchard, L.I. and Boyle, D.B. (2010) Detection and quantitation of gallid herpesvirus 1 in avian samples by 5′ taq nuclease assay utilizing minor groove binder technology. Avian Pathology, 39 (1). pp. 47-52. ISSN 03079457 (ISSN)

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Article Link(s): https://doi.org/10.1080/03079450903473582

Abstract

A 5′ Taq nuclease assay utilizing Minor Groove Binder technology and targeting the thymidine kinase gene of gallid herpesvirus 1 (GaHV-1) was designed and optimized for use in diagnosing avian infectious laryngotracheitis. The assay was specific for GaHV-1 in that it did not react with other avian viral or bacterial pathogens. The detection limit was 1.0-10-2 median tissue culture infectious dose per reaction or 90 target copies per reaction. Fifteen out of 41 diagnostic samples from sick birds reacted in the assay, five of which produced a typical alphaherpesvirus cytopathic effect (CPE) on chicken kidney (CK) cells. Sequencing, using amplicons generated by a polymerase chain reaction with primers flanking the 5′ Taq nuclease amplicon, confirmed the presence of GaHV-1 in six samples (two producing alphaherpesvirus CPE on CK cells, three not producing alphaherpesvirus CPE, and one that was not inoculated onto CK cells). Tracheal swabs taken from 18 healthy broilers did not react in the assay. The ability of the assay to determine viral load in samples was demonstrated. Overall the assay is suitable for the rapid diagnosis of infectious laryngotracheitis. © 2010 Houghton Trust Ltd.

Item Type:Article
Business groups:Animal Science, Biosecurity Queensland
Keywords:thymidine kinase virus DNA animal animal disease article bioassay bird disease chicken cytology cytopathogenic effect diagnosis, measurement and analysis genetics herpes virus infection Iltovirus isolation and purification kidney laryngitis methodology reverse transcription polymerase chain reaction trachea tracheitis virology Animals Biological Assay Chickens Cytopathogenic Effect, Viral DNA, Viral Herpesviridae Infections Herpesvirus 1, Gallid Laboratory Techniques and Procedures Poultry Diseases Reverse Transcriptase Polymerase Chain Reaction Aves Bacteria (microorganisms) Gallid herpesvirus 1
Subjects:Animal culture > Poultry
Veterinary medicine > Veterinary virology
Veterinary medicine > Veterinary pathology
Deposited On:17 Jan 2023 03:55
Last Modified:17 Jan 2023 03:55

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