RPA-CRISPR-Cas13a-assisted detection method of transmissible gastroenteritis virus Article Swipe
Haoyu Wang
,
Zhimeng Cheng
,
Ran Luo
,
Qiyue Yang
,
Yongping Zeng
,
Yijun Yang
,
Yuankun Chen
,
Wenting Li
,
Xiao Liu
·
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.3389/fvets.2024.1428591
YOU?
·
· 2024
· Open Access
·
· DOI: https://doi.org/10.3389/fvets.2024.1428591
Background and aim Transmissible gastroenteritis virus (TGEV) is a highly contagious gastrointestinal virus that causes diarrhea, vomiting, anorexia, dehydration, and weight loss in piglets. In clinical practice, it often occurs in mixed infections with other pathogens, and is therefore difficult to diagnose and prevent. It mainly harms piglets of about 2 weeks old, causing huge losses on farms. The clinical confirmation of TGEV usually requires a laboratory diagnosis, but traditional PCR and immunofluorescence assays have some limitations. Moreover, most farms in China are ill-equipped to accurately diagnose the disease. Therefore, a new detection method with high sensitivity and specificity and less dependence on instrumentation is required. Methods We used recombinase polymerase amplification (RPA), combined with the nuclease characteristics of the activated Cas13a protein to establish a visual CRISPR-Cas13a-assisted detection method for TGEV by adding a reporter RNA with fluorescent and quenching moieties to the system. Result We selected the optimal RPA primer and best CRISPR RNA (crRNA). The reaction system was optimized and its repeatability, specificity, and sensitivity verified. The TGEV detection system did not cross-react with other common diarrhea viruses, and its detection limit was 10 1 copies, which is similar with the sensitivity of qPCR. We successfully established an RPA–CRISPR-Cas13a-assisted detection method, and used this detection system to analyze 123 pig blood samples. qPCR was used as the gold standard method. The sensitivity, specificity, positive coincidence rate, and negative coincidence rate of the new method were 100, 98.93, 96.66, and 100%, respectively.
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.3389/fvets.2024.1428591
- OA Status
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- Cited By
- 2
- References
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- Related Works
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- OpenAlex ID
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All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4400261509Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.3389/fvets.2024.1428591Digital Object Identifier
- Title
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RPA-CRISPR-Cas13a-assisted detection method of transmissible gastroenteritis virusWork title
- Type
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articleOpenAlex work type
- Language
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enPrimary language
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2024Year of publication
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2024-07-02Full publication date if available
- Authors
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Haoyu Wang, Zhimeng Cheng, Ran Luo, Qiyue Yang, Yongping Zeng, Yijun Yang, Yuankun Chen, Wenting Li, Xiao LiuList of authors in order
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https://doi.org/10.3389/fvets.2024.1428591Publisher landing page
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YesWhether a free full text is available
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goldOpen access status per OpenAlex
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https://doi.org/10.3389/fvets.2024.1428591Direct OA link when available
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CRISPR, Virology, Biology, Genetics, GeneTop concepts (fields/topics) attached by OpenAlex
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2Total citation count in OpenAlex
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2025: 1, 2024: 1Per-year citation counts (last 5 years)
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