
A real-time fluorescent RT-PCR assay was developed to amplify avian paramyxovirus serotype 1 (APMV-1)-specific nucleic acid fragments from field samples. Subsequent 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 RT-PCR test for detection of a range of field cases and reference strains of APMV-1. Subsequent restriction endonuclease analysis 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 2004, seven cases of pigeon PMV-1 in Northern Ireland were diagnosed and differentiated more rapidly using the fluorescent RT-PCR assay when compared with the use of virus isolation. We report the development and application of a one-step real-time RT-PCR test coupled with REA as a fast, specific method for both the detection and typing of APMV-1 from field samples.
Virulence, Reverse Transcriptase Polymerase Chain Reaction, Newcastle Disease, Newcastle disease virus, Animals, Reproducibility of Results, Chickens, Sensitivity and Specificity
Virulence, Reverse Transcriptase Polymerase Chain Reaction, Newcastle Disease, Newcastle disease virus, Animals, Reproducibility of Results, Chickens, Sensitivity and Specificity
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