Rapid and visual detection of Lawsonia intracellularis with an improved recombinase polymerase amplification assay combined with a lateral flow dipstick.
BMC Vet Res
; 15(1): 97, 2019 Mar 21.
Article
em En
| MEDLINE
| ID: mdl-30898117
ABSTRACT
BACKGROUND:
Lawsonia intracellularis (L. intracellularis) is the etiologic agent of porcine proliferative enteropathy (PPE), which is reported in many swine breeding countries all over the world, and has caused enormous economic losses in intensive pig production systems. Therefore, the aim of this study was to develop a simple and rapid method for on-site detection of Lawsonia intracellularis (L. intracellularis). As the isothermal recombinase polymerase amplification (RPA) can be performed at a constant temperature and its product is directly observed on a lateral-flow dipstick (LFD) with naked eyes without electrophoresis, the RPA-LFD assay should be useful for field diagnosis of L. intracellularis as well as its detection from clinical samples.RESULTS:
The established RPA-LFD assay could be finished in 30 min at a wide temperature range of 25 to 40 °C, and the amplicons could be visualized by naked eyes. The developed RPA-LFD assay was specific to dnaA gene of L. intracellularis, and did not detect nucleic acids extracted from other common gastrointestinal pathogens. The minimum detection of this RPA-LFD method was 400 L. intracellularis per reaction, which was as sensitive as conventional PCR. Further, the RPA-LFD assay was performed with 150 clinical fecal samples and the detection results were compared with conventional PCR. Results showed that the coincidence rate of RPA-LFD and conventional PCR was 100%.CONCLUSIONS:
The combined RPA with LFD assay provides a simple, rapid, specific and sensitive alternative for field diagnosis of L. intracellularis infection.Palavras-chave
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Kit de Reagentes para Diagnóstico
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Doenças dos Suínos
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Lawsonia (Bactéria)
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Técnicas de Amplificação de Ácido Nucleico
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Técnicas de Diagnóstico Molecular
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Infecções por Desulfovibrionaceae
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
Idioma:
En
Revista:
BMC Vet Res
Assunto da revista:
MEDICINA VETERINARIA
Ano de publicação:
2019
Tipo de documento:
Article
País de afiliação:
China