A rapid RT-LAMP assay for the detection of all four lineages of Peste des Petits Ruminants Virus.
J Virol Methods
; 274: 113730, 2019 12.
Article
en En
| MEDLINE
| ID: mdl-31513860
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 < 20â¯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 TCID50â¯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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Virus de la Peste de los Pequeños Rumiantes
/
Técnicas de Amplificación de Ácido Nucleico
/
Peste de los Pequeños Rumiantes
/
Técnicas de Diagnóstico Molecular
Tipo de estudio:
Diagnostic_studies
/
Evaluation_studies
Límite:
Animals
Idioma:
En
Revista:
J Virol Methods
Año:
2019
Tipo del documento:
Article
Pais de publicación:
HOLANDA
/
HOLLAND
/
NETHERLANDS
/
NL
/
PAISES BAJOS
/
THE NETHERLANDS