A rapid on-site visualization platform based on RPA coupled with CRISPR-Cas12a for the detection of genetically modified papaya ‘Huanong No.1’

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...

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Published in:	Talanta (Oxford) Vol. 277; p. 126437
Main Authors:	Zhu, Lili, He, Gongwen, Yang, Guiqin, Yang, Wenli, He, Ying, Chen, Jian, Chen, Yanxin, Ji, Yi, Pan, Zhiwen, Yao, Juan, Chen, Xiaoyun, Jiang, Dagang
Format:	Journal Article
Language:	English
Published:	Netherlands Elsevier B.V 01.09.2024
Subjects:	China CRISPR-Cas12a detection limit GM papaya Huanong No.1 Papaya ringspot virus rapid methods recombinase polymerase amplification RPA Visualization China Visualization CRISPR-Cas12a RPA GM papaya Huanong No.1
ISSN:	0039-9140, 1873-3573, 1873-3573
Online Access:	Get full text
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Abstract	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. •The RPA-CRISPR-Cas12a platform was first developed for the detection of genetically modified papaya ‘Huanong No.1’.•This novel platform enabled the on-site visualization detection of ‘Huanong No.1’, eliminating dependence on laboratory conditions and specialized instruments.•The detection process was faster than other detection methods and could be completed in less than 1 h.•The developed RPA-CRISPR-Cas12a platform showed highly specificity and sensitivity for ‘Huanong No.1’ detection.
AbstractList	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.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. 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. 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. 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. •The RPA-CRISPR-Cas12a platform was first developed for the detection of genetically modified papaya ‘Huanong No.1’.•This novel platform enabled the on-site visualization detection of ‘Huanong No.1’, eliminating dependence on laboratory conditions and specialized instruments.•The detection process was faster than other detection methods and could be completed in less than 1 h.•The developed RPA-CRISPR-Cas12a platform showed highly specificity and sensitivity for ‘Huanong No.1’ detection.
ArticleNumber	126437
Author	Ji, Yi Chen, Yanxin Yang, Wenli Chen, Xiaoyun Yang, Guiqin He, Ying Yao, Juan Jiang, Dagang He, Gongwen Zhu, Lili Chen, Jian Pan, Zhiwen
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Keywords	Visualization CRISPR-Cas12a RPA GM papaya Huanong No.1
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Snippet	The Papaya ringspot virus (PRSV)-resistant genetically modified (GM) papaya ‘Huanong No.1’ has been certified as safe for consumption and widely planted in... The Papaya ringspot virus (PRSV)-resistant genetically modified (GM) papaya 'Huanong No.1' has been certified as safe for consumption and widely planted in...
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SubjectTerms	China CRISPR-Cas12a detection limit GM papaya Huanong No.1 Papaya ringspot virus rapid methods recombinase polymerase amplification RPA Visualization
Title	A rapid on-site visualization platform based on RPA coupled with CRISPR-Cas12a for the detection of genetically modified papaya ‘Huanong No.1’
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