Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 265, Issue -, Pages 9-14Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2018.12.001
Keywords
Newcastle disease; Avian avulavirus 1; rRT-PCR; MGB probe
Funding
- UK Department for Environment, Food and Rural Affairs (Defra)
- devolved government (Wales) [SE2205, SE2208]
- devolved government (Scotland) [SE2205, SE2208]
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Newcastle disease is a devastating disease of poultry caused by Newcastle disease virus (NDV), a virulent form of avian avulavirus 1 (AAvV-1). A rapid, sensitive and specific means for the detection of NDV is fundamental for the control of this notifiable transboundary virus. Although several real-time RT-PCR assays exist for the detection of AAvV-1, diagnostic sensitivity and specificities can be sub-optimal. In this study, we describe a modification to an existing AAvV-1 L-gene RT-PCR screening assay, where the original probe set was replaced with minor groove binding (MGB) probes, to create the MGB L-gene assay. The diagnostic sensitivity and specificity of this assay was evaluated against a broad panel of both Class I and Class II AAvV-1 viruses of diverse and representative lineages/genotypes in both clinical samples and amplified viruses, and compared with a number of previously published real-time RT-PCR screening assays for AAvV-1. The MGB L-gene assay out-performed all other assays in this assessment, with enhanced sensitivity and specificity, detecting isolates from a broad range of virus lineages/genotypes (including contemporaneously-circulating strains). The assay has also proved its value for screening original clinical samples for the presence of AAvV-1, thus providing an improved screening assay for routine detection of this notifiable disease agent.
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