Development of an L gene real-time reverse-transcription PCR assay for the detection of avian paramyxovirus type 1 RNA in clinical samples

• Published in: Archives of virology Vol. 155; no. 6; pp. 817 - 823
• Main Authors: Fuller, Chad M, Brodd, Lina, Irvine, Richard M, Alexander, Dennis J, Aldous, Elizabeth W
• Format: Journal Article
• Language: English
• Published: Vienna Vienna : Springer Vienna 01.06.2010; Springer Vienna; Springer; Springer Nature B.V
• Subjects: Animals; Biological and medical sciences; Biomedical and Life Sciences; Biomedicine; Bird Diseases - diagnosis; Bird Diseases - virology; Birds; Chickens; Fundamental and applied biological sciences. Psychology; Infectious Diseases; Medical Microbiology; Microbiology; Miscellaneous; Newcastle Disease - diagnosis; Newcastle Disease - virology; Newcastle disease virus; Newcastle disease virus - genetics; Newcastle disease virus - isolation & purification; Oligonucleotide Probes; Original Article; Paramyxovirus; Poultry Diseases - diagnosis; Poultry Diseases - virology; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction - methods; Reverse Transcriptase Polymerase Chain Reaction - veterinary; Sensitivity and Specificity; Viral Proteins - genetics; Virology; Grey Partridge; Avian Influenza; Infectious Bronchitis Virus; Newcastle Disease Virus; Domestic Poultry; Virus; Paramyxovirinae; Mononegavirales; Gene; Rubulavirus; Newcastle disease virus; Detection; Real time; Veterinary; Reverse transcription polymerase chain reaction; Paramyxoviridae
• ISSN: 0304-8608; 1432-8798
• DOI: 10.1007/s00705-010-0632-1

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.