Differentiation of virulent and non-virulent strains of Newcastle disease virus within 24 hours by polymerase chain reaction

Fast diagnosis of Newcastle disease is a prerequisite for confining outbreaks. Diagnosis implies the differentiation of virulent and non-virulent Newcastle disease viruses (NDV). However, conventional methods, i.e. isolation of the virus and determination of the intracerebral pathogenicity index, ta...

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Bibliographic Details
Published in:Avian pathology 1997, Vol.26 (4), p.837-849
Main Authors: Kant, A, Koch, G, Roozelaar, D.J. van, Balk, F, Huurne, A. ter
Format: Article
Language:English
Subjects:
differential diagnosis
ID Lelystad, Institute for Animal Science and Health
ID-Lelystad, Instituut voor Dierhouderij en Diergezondheid
Newcastle disease virus
Pathology
polymerase chain reaction
Poultry
rapid methods
Ribonucleic acid
RNA
RNA-directed DNA polymerase
strain differences
Studies
virulence
Viruses
Wildfowl
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Summary:Fast diagnosis of Newcastle disease is a prerequisite for confining outbreaks. Diagnosis implies the differentiation of virulent and non-virulent Newcastle disease viruses (NDV). However, conventional methods, i.e. isolation of the virus and determination of the intracerebral pathogenicity index, take at least 5 days. Therefore, we investigated whether diagnosis can be performed by using the reverse transcriptase-polymerase chain reaction (RT-PCR) on RNA isolated directly from tissue homogenate. Two oligonucleotide primers, representing the sequence at the cleavage site of the F protein of either virulent or non-virulent NDV strains, respectively, were used to differentiate NDV. Using the RT-PCR we were able to differentiate 15 NDV reference strains, 11 of which were virulent and 14 non-virulent. The RT-PCR was further validated by using homogenate of brain, trachea, lung and spleen from 12 chicken flocks and one turkey flock suspected of Newcastle disease. The RT-PCR detected virulent NDV in samples of seven flocks and non-virulent NDV in two out of three flocks in agreement with conventional methods. However the RT-PCR failed to detect virus in 1/3 flocks from which non-virulent virus was isolated. The results are discussed. We conclude that the RT-PCR described can be used to confirm diagnosis of Newcastle disease within 24 h using RNA isolated directly from tissue homogenate.
ISSN:0307-9457
1465-3338
DOI:10.1080/03079459708419257