Development of a reverse transcription-loop-mediated isothermal amplification assay for the detection of parrot bornavirus 4.
J Virol Methods
; 275: 113749, 2020 01.
Article
en En
| MEDLINE
| ID: mdl-31622637
ABSTRACT
Avian bornavirus (ABV) is the causative agent of proventricular dilatation disease, which is fatal in psittacine birds. ABVs have spread worldwide, and outbreaks have led to mass deaths of captive birds in commercial and breeding facilities. The segregation of infected birds is a countermeasure to prevent ABV spread in aviaries. However, this approach requires a highly sensitive detection method for the screening of infected birds before virus transmission. In this study, we developed a reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay for the diagnosis of parrot bornavirus 4 (PaBV-4), a dominant ABV genotype. Using this assay, we successfully detected PaBV-4 RNA in cell cultures, brain tissues, and feces. We also developed methods for simple RNA extraction and visual detection without electrophoresis. The sensitivity of the newly established RT-LAMP assay was 100-fold higher than that of the real-time PCR (RT-qPCR) assay. Accordingly, the RT-LAMP assay developed in this study is suitable for the rapid and sensitive diagnosis of PaBV-4 without specialized equipment and will contribute to virus control in aviaries.
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Base de datos:
MEDLINE
Asunto principal:
Loros
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Enfermedades de las Aves
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Infecciones por Mononegavirales
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Técnicas de Amplificación de Ácido Nucleico
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Bornaviridae
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Técnicas de Diagnóstico Molecular
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Transcripción Reversa
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
J Virol Methods
Año:
2020
Tipo del documento:
Article