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Journal of Clinical Microbiology, September 2001, p. 3171-3178, Vol. 39, No. 9
Instituto de Biotecnologia, Centro de Investigacion en
Ciencias Veterinarias, Instituto Nacional de Tecnologia
Agropecuria, CC7725 Castelar (1712), Buenos
Aires,1 and CONICET, Rivadavia 1917 (1033), Capital Federal,2 Argentina, and
Southeast Poultry Research Laboratory, USDA Agricultural
Research Service, Athens, Georgia 306053
Received 19 March 2001/Returned for modification 17 June
2001/Accepted 1 July 2001
Newcastle disease virus (NDV) is an economically important pathogen
of poultry that may cause clinical disease that ranges from a mild
respiratory syndrome to a virulent form with high mortality, depending
on an isolate's pathotype. Infections with virulent NDV strains are
required to be reported by member nations to the Office of
International Epizootes (OIE). The primary determinant for virulence
among NDV isolates is the presence or absence of dibasic amino acids in
the fusion (F) protein cleavage activation site. Along with biological
virulence determinations as the definitive tests, OIE accepts reporting
of the F protein cleavage site sequence of NDV isolates as a virulence
criterion. Nucleotide sequence data for many NDV isolates recently
isolated from infected chickens and other avian species worldwide have
been deposited in GenBank. Consequently, viral genomic information
surrounding the F protein cleavage site coding sequence was used to
develop a heteroduplex mobility assay (HMA) to aid in further
identification of molecular markers as predictors of NDV virulence.
Using common vaccine strains as a reference, we were able to
distinguish virulent viruses among NDV isolates that correlated with
phylogenetic analysis of the nucleotide sequence. This technique was
also used to examine NDV isolates not previously characterized. We were
able to distinguish vaccine-like viruses from other isolates
potentially virulent for chickens. This technique will help improve
international harmonization of veterinary biologics as set forth by the
OIE and the Veterinary International Cooperation on Harmonization of
Technical Requirements of Veterinary Medicinal Products. Ultimately,
the HMA could be used for initial screening among a large number of
isolates and rapid identification of potentially virulent NDV that
continue to threaten commercial poultry worldwide.
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.9.3171-3178.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Use of a Heteroduplex Mobility Assay To Detect
Differences in the Fusion Protein Cleavage Site Coding Sequence
among Newcastle Disease Virus Isolates

*
Corresponding author. Mailing address: Southeast
Poultry Research Laboratory, A.R.S., U.S.D.A., 934 College Station Rd.,
Athens, GA 30605. Phone: (706) 546-3463 or (706) 546-3434. Fax: (706) 546-3161. E-mail: bseal{at}seprl.usda.gov.
Present address: Department of Avian Medicine, College of
Veterinary Medicine, University of Georgia, Athens, GA 30602.
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