Rapid detection of duck hepatitis A virus genotype C using reverse transcription loop-mediated isothermal amplification.
J Virol Methods
; 196: 193-8, 2014 Feb.
Article
en En
| MEDLINE
| ID: mdl-24291148
A one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was used and optimized to develop a rapid and sensitive detection system for duck hepatitis A virus genotype C (DHAV-C) RNA. A set of four specific primers was designed against highly conserved sequences located within the 3D gene from DHAV (strain GX1201). Under optimal reaction conditions, the sensitivity of DHAV-C-specific RT-LAMP was 100-fold higher than that of reverse transcriptase-polymerase chain reaction (RT-PCR), with a detection limit of 0.3pg (6.59×10(4) copies) per reaction. No cross-reactivity was observed from the samples of other duck viruses, which is in good accordance with RT-PCR. Furthermore, a positive reaction can be visually inspected by observing turbidity or color change after the addition of SYBR green I dye. The DHAV-C-specific RT-LAMP assay was applied to the samples and compared with RT-PCR. The positive-sample ratios were 26.7% (12 of 45) by RT-LAMP and 20% (9 of 45) by RT-PCR. Therefore, the newly developed RT-LAMP assay is a rapid, specific, sensitive, and cost-effective method of DHAV-C detection. This assay has potential applications in both clinical diagnosis and field surveillance of DHAV-C infection.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Enfermedades de las Aves de Corral
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Medicina Veterinaria
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Virología
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Infecciones por Picornaviridae
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Técnicas de Amplificación de Ácido Nucleico
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Técnicas de Diagnóstico Molecular
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Virus de la Hepatitis del Pato
Tipo de estudio:
Diagnostic_studies
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Evaluation_studies
Límite:
Animals
Idioma:
En
Revista:
J Virol Methods
Año:
2014
Tipo del documento:
Article
País de afiliación:
China
Pais de publicación:
Países Bajos