CRISPR-Cas13a-based detection method for avian influenza virus
Avian influenza virus (AIV) causes huge losses to the global poultry industry and poses a threat to humans and other mammals. Fast, sensitive, and portable diagnostic methods are essential for efficient avian influenza control. Here, a clustered regularly interspaced short palindromic repeats (CRISP...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-10-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1288951/full |
| _version_ | 1797662097607229440 |
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| author | Yuhan Wu Jiaxing Zhan Zhaomeng Shan Yanbing Li Yining Liu Yan Li Yixin Wang Zhe Liu Xuexia Wen Xiurong Wang |
| author_facet | Yuhan Wu Jiaxing Zhan Zhaomeng Shan Yanbing Li Yining Liu Yan Li Yixin Wang Zhe Liu Xuexia Wen Xiurong Wang |
| author_sort | Yuhan Wu |
| collection | DOAJ |
| description | Avian influenza virus (AIV) causes huge losses to the global poultry industry and poses a threat to humans and other mammals. Fast, sensitive, and portable diagnostic methods are essential for efficient avian influenza control. Here, a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas13a based platform was developed to detect AIV. This novel method was developed to specifically detect H1–H16 subtypes of AIV with fluorescence and lateral flow-based readouts and exhibited no cross-reactivity with Newcastle disease virus, avian infectious bronchitis virus, or infectious bursal disease virus. The limit of detection was determined to be 69 and 690 copies/μL using fluorescence and lateral flow as readouts, respectively. The developed assay exhibited 100% consistency with quantitative real-time polymerase chain reaction in detecting clinical samples. The heating of unextracted diagnostic samples to obliterate nuclease treatment was introduced to detect viral RNA without nucleic acid extraction. Single-step optimization was used to perform reverse transcription, recombinase polymerase amplification, and CRISPR-Cas13a detection in a tube. These advances resulted in an optimized assay that could specifically detect AIV with simplified procedures and reduced contamination risk, highlighting the potential to be used in point-of-care testing. |
| first_indexed | 2024-03-11T18:55:10Z |
| format | Article |
| id | doaj.art-05042f01389941dea0d0f5624fb00dbd |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-03-11T18:55:10Z |
| publishDate | 2023-10-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-05042f01389941dea0d0f5624fb00dbd2023-10-11T05:05:27ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2023-10-011410.3389/fmicb.2023.12889511288951CRISPR-Cas13a-based detection method for avian influenza virusYuhan Wu0Jiaxing Zhan1Zhaomeng Shan2Yanbing Li3Yining Liu4Yan Li5Yixin Wang6Zhe Liu7Xuexia Wen8Xiurong Wang9Key Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, ChinaKey Laboratory of Livestock Infectious Diseases, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, ChinaState Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin, ChinaAvian influenza virus (AIV) causes huge losses to the global poultry industry and poses a threat to humans and other mammals. Fast, sensitive, and portable diagnostic methods are essential for efficient avian influenza control. Here, a clustered regularly interspaced short palindromic repeats (CRISPR)-Cas13a based platform was developed to detect AIV. This novel method was developed to specifically detect H1–H16 subtypes of AIV with fluorescence and lateral flow-based readouts and exhibited no cross-reactivity with Newcastle disease virus, avian infectious bronchitis virus, or infectious bursal disease virus. The limit of detection was determined to be 69 and 690 copies/μL using fluorescence and lateral flow as readouts, respectively. The developed assay exhibited 100% consistency with quantitative real-time polymerase chain reaction in detecting clinical samples. The heating of unextracted diagnostic samples to obliterate nuclease treatment was introduced to detect viral RNA without nucleic acid extraction. Single-step optimization was used to perform reverse transcription, recombinase polymerase amplification, and CRISPR-Cas13a detection in a tube. These advances resulted in an optimized assay that could specifically detect AIV with simplified procedures and reduced contamination risk, highlighting the potential to be used in point-of-care testing.https://www.frontiersin.org/articles/10.3389/fmicb.2023.1288951/fullavian influenza virusrecombinase polymerase amplificationCRISPR-cas13alateral flow dipstick assayuniversal detection |
| spellingShingle | Yuhan Wu Jiaxing Zhan Zhaomeng Shan Yanbing Li Yining Liu Yan Li Yixin Wang Zhe Liu Xuexia Wen Xiurong Wang CRISPR-Cas13a-based detection method for avian influenza virus Frontiers in Microbiology avian influenza virus recombinase polymerase amplification CRISPR-cas13a lateral flow dipstick assay universal detection |
| title | CRISPR-Cas13a-based detection method for avian influenza virus |
| title_full | CRISPR-Cas13a-based detection method for avian influenza virus |
| title_fullStr | CRISPR-Cas13a-based detection method for avian influenza virus |
| title_full_unstemmed | CRISPR-Cas13a-based detection method for avian influenza virus |
| title_short | CRISPR-Cas13a-based detection method for avian influenza virus |
| title_sort | crispr cas13a based detection method for avian influenza virus |
| topic | avian influenza virus recombinase polymerase amplification CRISPR-cas13a lateral flow dipstick assay universal detection |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2023.1288951/full |
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