CRISPR/Cas13a-based genome editing for establishing the detection method of H9N2 subtype avian influenza virus.
Poult Sci
; 103(10): 104068, 2024 Oct.
Article
en En
| MEDLINE
| ID: mdl-39096825
ABSTRACT
Avian influenza virus (AIV) subtype H9N2 has significantly threatened the poultry business in recent years by having become the predominant subtype in flocks of chickens, ducks, and pigeons. In addition, the public health aspects of H9N2 AIV pose a significant threat to humans. Early and rapid diagnosis of H9N2 AIV is therefore of great importance. In this study, a new method for the detection of H9N2 AIV based on fluorescence intensity was successfully established using CRISPR/Cas13a technology. The Cas13a protein was first expressed in a prokaryotic system and purified using nickel ion affinity chromatography, resulting in a high-purity Cas13a protein. The best RPA (recombinase polymerase amplification) primer pairs and crRNA were designed and screened, successfully constructing the detection of H9N2 AIV based on CRISPR/Cas13a technology. Optimal concentration of Cas13a and crRNA was determined to optimize the constructed assay. The sensitivity of the optimized detection system is excellent, with a minimum detection limit of 10° copies/µL and didn't react with other avian susceptible viruses, with excellent specificity. The detection method provides the basis for the field detection of the H9N2 AIV.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Enfermedades de las Aves de Corral
/
Pollos
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Subtipo H9N2 del Virus de la Influenza A
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Sistemas CRISPR-Cas
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Edición Génica
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Gripe Aviar
Límite:
Animals
Idioma:
En
Revista:
Poult Sci
Año:
2024
Tipo del documento:
Article
Pais de publicación:
Reino Unido