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Talanta ; 277: 126437, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38901194

RESUMO

The Papaya ringspot virus (PRSV)-resistant genetically modified (GM) papaya 'Huanong No.1' has been certified as safe for consumption and widely planted in China for about 18 years. To protect consumers' rights and facilitate government supervision and monitoring, it is necessary to establish a simple, rapid, and specific detection method for 'Huanong No.1'. Herein, we developed a platform based on recombinase polymerase amplification (RPA) coupled with CRISPR-Cas12a for the detection of 'Huanong No.1'. The RPA-CRISPR-Cas12a platform was found to have high specificity, with amplification signals only present in 'Huanong No.1'. Additionally, the platform was highly sensitive, with a limit of detection (LOD) of approximately 20 copies. The detection process was fast and could be completed in less than 1 h. This novel platform enables the rapid on-site visualization detection of 'Huanong No.1', eliminating dependence on laboratory conditions and specialized instruments, and can serve as a technical reference for the rapid detection of other GM plants.


Assuntos
Sistemas CRISPR-Cas , Carica , Técnicas de Amplificação de Ácido Nucleico , Plantas Geneticamente Modificadas , Carica/genética , Carica/virologia , Sistemas CRISPR-Cas/genética , Plantas Geneticamente Modificadas/genética , Técnicas de Amplificação de Ácido Nucleico/métodos , Potyvirus/genética , Potyvirus/isolamento & purificação , Recombinases/metabolismo , Limite de Detecção , Proteínas de Bactérias , Endodesoxirribonucleases , Proteínas Associadas a CRISPR
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