Development of an L gene real-time reverse-transcription PCR assay for the detection of avian paramyxovirus type 1 RNA in clinical samples.
A real-time reverse-transcription PCR (rRT-PCR) that targets a region of the polymerase (L) gene was developed to detect all known lineages of avian paramyxovirus type 1 (APMV-1), also known as Newcastle disease virus (NDV). A panel of 23 viruses representing the current known phylogenetic diversity of the APMV-1 population with a bias towards the more recent European strains, which had been grown in embryonated fowls' eggs, were tested. A range of positive and negative clinical samples (n = 350) provided by the National Reference Laboratory and International Reference Laboratory at VLA Weybridge were also tested. Positive clinical material included samples considered representative of lineages 3, 4 and 5 obtained from chickens, ducks, pigeons and partridges. The negative sample population was obtained from chickens, turkeys and ducks. The APMV-1 L gene rRT-PCR gave high relative sensitivity (96.05%) and specificity (98.18%) when compared with virus isolation in embryonated fowls' eggs. It is proposed that this assay could provide a first-line screening tool for the detection of APMV-1 in clinical samples.
Fuller
Chad M
CM
Veterinary Laboratories Agency, Surrey, UK. c.fuller@vla.defra.gsi.gov.uk
Brodd
Lina
L
Irvine
Richard M
RM
Alexander
Dennis J
DJ
Aldous
Elizabeth W
EW
D000818:Animals; D001715:Bird Diseases / Q000175:diagnosis / Q000821:virology; D001717:Birds; D002645:Chickens; D009521:Newcastle Disease / Q000175:diagnosis* / Q000821:virology*; D009522:Newcastle disease virus / Q000235:genetics / Q000302:isolation & purification*; D015345:Oligonucleotide Probes; D011201:Poultry Diseases / Q000175:diagnosis / Q000821:virology; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary; D012680:Sensitivity and Specificity; D014764:Viral Proteins / Q000235:genetics*
C431770:L protein, paramyxoviridae; D015345:Oligonucleotide Probes; D014764:Viral Proteins