Detection and differentiation of pathogenicity of avian paramyxovirus serotype 1 from field cases using one-step reverse transcriptase-polymerase chain reaction.
Amplification of avian paramyxovirus serotype 1 (APMV-1)-specific nucleic acid fragments, followed by restriction endonuclease analysis (REA) using BglI, was carried out to type strains according to their virulence. Primer sequences were used to amplify a 202 base pair fragment, encompassing the fusion protein cleavage site, in a one-step reverse transcriptase-polymerase chain reaction (RT-PCR) test for detection of a range of field cases and reference strains of APMV-1. Subsequent REA of the amplified fragments enabled differentiation of low virulent lentogenic field and vaccine strains from more virulent mesogenic and velogenic field strains of APMV-1, including pigeon PMV-1. In the present paper, we report the development and application of a one-step RT-PCR test coupled with REA as a fast, specific method for both the detection and typing of APMV-1 from field samples.
Creelan
Julie L
JL
Veterinary Science Division, Department of Agriculture and Rural Development, Stormont, Belfast BT4 3SD, Northern Ireland, UK. Julie.Creelan@dardni.gov.uk
Graham
David A
DA
McCullough
Samuel J
SJ
D000818:Animals; D002645:Chickens; D010856:Columbidae; D009521:Newcastle Disease / Q000473:pathology*; D009522:Newcastle disease virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification / Q000472:pathogenicity*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D011201:Poultry Diseases / Q000473:pathology* / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012703:Serotyping / Q000662:veterinary; D014774:Virulence