A rapid RT-LAMP assay for the detection of all four lineages of Peste des Petits Ruminants Virus.

Abstract:

:Peste des petits ruminants (PPR) is a viral disease of small ruminants that is caused by the PPR virus (PPRV) and is a significant burden on subsistence farmers across the developing world. Loop-mediated isothermal amplification (LAMP) provides cost-effective, rapid, specific and sensitive detection of nucleic acid and has been demonstrated to have field application for a range of viruses. We describe the development of a novel PPRV RT-LAMP assay utilising carefully-selected primers (targeting the N-gene) allowing for the detection of all known PPRV lineages in < 20 min. The assay was evaluated in comparison with a "gold standard" real-time RT-PCR assay using more than 200 samples, comprising samples from recent PPRV outbreaks, experimentally-infected goats, well-characterised cell culture isolates and samples collected from uninfected animals. The RT-LAMP assay demonstrated 100% diagnostic specificity and greater than 97% diagnostic sensitivity in comparison with the real-time RT-PCR assay. The limit of detection was between 0.3 and 0.8 log10 TCID50 ml-1 equating to a CT value of 31.52 to 33.48. In experimentally-infected animals, the RT-LAMP could detect PPRV as early as 4 days post infection (dpi) - before clinical signs were observed at 7 dpi. The RT-LAMP assay can support the global PPR eradication campaign.

journal_name

J Virol Methods

authors

Rajko-Nenow P,Flannery J,Arnold H,Howson ELA,Darpel K,Stedman A,Corla A,Batten C

doi

10.1016/j.jviromet.2019.113730

subject

Has Abstract

pub_date

2019-12-01 00:00:00

pages

113730

eissn

0166-0934

issn

1879-0984

pii

S0166-0934(19)30298-8

journal_volume

274

pub_type

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