Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay
IntroductionSenecavirus A (SVA) is a highly contagious virus that causes vesicular disease in pigs. At present, laboratory detection methods, such as virus isolation and polymerase chain reaction (PCR), required precision instruments and qualified personnel, making them unsuitable for point-of-care...
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1305222/full |
| _version_ | 1797362295019405312 |
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| author | Lei Ma Lei Ma Mengjie Zhu Qingfeng Meng Yao Wang Xueping Wang Xueping Wang |
| author_facet | Lei Ma Lei Ma Mengjie Zhu Qingfeng Meng Yao Wang Xueping Wang Xueping Wang |
| author_sort | Lei Ma |
| collection | DOAJ |
| description | IntroductionSenecavirus A (SVA) is a highly contagious virus that causes vesicular disease in pigs. At present, laboratory detection methods, such as virus isolation and polymerase chain reaction (PCR), required precision instruments and qualified personnel, making them unsuitable for point-of-care tests (POCT). Fortunately, the emergence of CRISPR/Cas system has provided new opportunities for fast and efficient pathogen detection.MethodsThis study successfully developed a precise and sensitive detection platform for diagnosing SVA by combining the CRISPR system with recombinase polymerase amplification (RPA). ResultsThe minimum detection limit of the assay was 10 copies of the SVA genome. Meanwhile, the assay demonstrated high specificity. To validate the effectiveness of this system, we tested 85 swine clinical samples and found that the fluorescence method had a 100% coincidence rate compared to RT-qPCR. DiscussionOverall, the RPA-CRISPR/Cas12a assay established in our study is a highly effective method for detecting SVA and holds great potential for practical applications in the resource-limited settings. |
| first_indexed | 2024-03-08T16:06:13Z |
| format | Article |
| id | doaj.art-288e66aae6c24cee81b46656189a2cff |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-03-08T16:06:13Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-288e66aae6c24cee81b46656189a2cff2024-01-08T05:38:31ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882024-01-011310.3389/fcimb.2023.13052221305222Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assayLei Ma0Lei Ma1Mengjie Zhu2Qingfeng Meng3Yao Wang4Xueping Wang5Xueping Wang6School of Biotechnology and Food Engineering, Anyang Institute of Technology, Anyang, ChinaCollege of Life Science, Henan University, Kaifeng, ChinaSchool of Biotechnology and Food Engineering, Anyang Institute of Technology, Anyang, ChinaTesting Technology R&D Department, Shanghai Kaiwosha Biotechnology Co., Ltd, Shanghai, ChinaSchool of Biotechnology and Food Engineering, Anyang Institute of Technology, Anyang, ChinaSchool of Biotechnology and Food Engineering, Anyang Institute of Technology, Anyang, ChinaCollege of Life Science, Henan University, Kaifeng, ChinaIntroductionSenecavirus A (SVA) is a highly contagious virus that causes vesicular disease in pigs. At present, laboratory detection methods, such as virus isolation and polymerase chain reaction (PCR), required precision instruments and qualified personnel, making them unsuitable for point-of-care tests (POCT). Fortunately, the emergence of CRISPR/Cas system has provided new opportunities for fast and efficient pathogen detection.MethodsThis study successfully developed a precise and sensitive detection platform for diagnosing SVA by combining the CRISPR system with recombinase polymerase amplification (RPA). ResultsThe minimum detection limit of the assay was 10 copies of the SVA genome. Meanwhile, the assay demonstrated high specificity. To validate the effectiveness of this system, we tested 85 swine clinical samples and found that the fluorescence method had a 100% coincidence rate compared to RT-qPCR. DiscussionOverall, the RPA-CRISPR/Cas12a assay established in our study is a highly effective method for detecting SVA and holds great potential for practical applications in the resource-limited settings.https://www.frontiersin.org/articles/10.3389/fcimb.2023.1305222/fullSenecavirus Arecombinase polymerase amplificationCRISPR/Cas12adetectionswine |
| spellingShingle | Lei Ma Lei Ma Mengjie Zhu Qingfeng Meng Yao Wang Xueping Wang Xueping Wang Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay Frontiers in Cellular and Infection Microbiology Senecavirus A recombinase polymerase amplification CRISPR/Cas12a detection swine |
| title | Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay |
| title_full | Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay |
| title_fullStr | Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay |
| title_full_unstemmed | Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay |
| title_short | Real-time detection of Seneca Valley virus by one-tube RPA-CRISPR/Cas12a assay |
| title_sort | real time detection of seneca valley virus by one tube rpa crispr cas12a assay |
| topic | Senecavirus A recombinase polymerase amplification CRISPR/Cas12a detection swine |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1305222/full |
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