Rapid and Accurate Campylobacter jejuni Detection With CRISPR-Cas12b Based on Newly Identified Campylobacter jejuni-Specific and -Conserved Genomic Signatures
Campylobacter jejuni is among the most prevalent foodborne zoonotic pathogens leading to diarrheal diseases. In this study, we developed a CRISPR-Cas12b-based system to rapidly and accurately detect C. jejuni contamination. Identification of C. jejuni-specific and -conserved genomic signatures is a...
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| Format: | Article |
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Frontiers Media S.A.
2021-04-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2021.649010/full |
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| author | Yu Huang Yu Huang Yu Huang Yu Huang Dan Gu Dan Gu Dan Gu Dan Gu Han Xue Han Xue Han Xue Han Xue Jinyan Yu Jinyan Yu Jinyan Yu Jinyan Yu Yuanyue Tang Yuanyue Tang Yuanyue Tang Yuanyue Tang Jinlin Huang Jinlin Huang Jinlin Huang Jinlin Huang Yunzeng Zhang Yunzeng Zhang Yunzeng Zhang Yunzeng Zhang Xinan Jiao Xinan Jiao Xinan Jiao Xinan Jiao |
| author_facet | Yu Huang Yu Huang Yu Huang Yu Huang Dan Gu Dan Gu Dan Gu Dan Gu Han Xue Han Xue Han Xue Han Xue Jinyan Yu Jinyan Yu Jinyan Yu Jinyan Yu Yuanyue Tang Yuanyue Tang Yuanyue Tang Yuanyue Tang Jinlin Huang Jinlin Huang Jinlin Huang Jinlin Huang Yunzeng Zhang Yunzeng Zhang Yunzeng Zhang Yunzeng Zhang Xinan Jiao Xinan Jiao Xinan Jiao Xinan Jiao |
| author_sort | Yu Huang |
| collection | DOAJ |
| description | Campylobacter jejuni is among the most prevalent foodborne zoonotic pathogens leading to diarrheal diseases. In this study, we developed a CRISPR-Cas12b-based system to rapidly and accurately detect C. jejuni contamination. Identification of C. jejuni-specific and -conserved genomic signatures is a fundamental step in development of the detection system. By comparing C. jejuni genome sequences with those of the closely related Campylobacter coli, followed by comprehensive online BLAST searches, a 20-bp C. jejuni-conserved (identical in 1024 out of 1037 analyzed C. jejuni genome sequences) and -specific (no identical sequence detected in non-C. jejuni strains) sequence was identified and the system was then assembled. In further experiments, strong green fluorescence was observed only when C. jejuni DNA was present in the system, highlighting the specificity of this system. The assay, with a sample-to-answer time of ∼40 min, positively detected chicken samples that were contaminated with a dose of approximately 10 CFU C. jejuni per gram of chicken, which was >10 times more sensitive than the traditional Campylobacter isolation method, suggesting that this method shows promise for onsite C. jejuni detection. This study provides an example of bioinformatics-guided CRISPR-Cas12b-based detection system development for rapid and accurate onsite pathogen detection. |
| first_indexed | 2024-12-17T22:26:43Z |
| format | Article |
| id | doaj.art-db34aa2939d44715a66825bad7786973 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-17T22:26:43Z |
| publishDate | 2021-04-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-db34aa2939d44715a66825bad77869732022-12-21T21:30:19ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2021-04-011210.3389/fmicb.2021.649010649010Rapid and Accurate Campylobacter jejuni Detection With CRISPR-Cas12b Based on Newly Identified Campylobacter jejuni-Specific and -Conserved Genomic SignaturesYu Huang0Yu Huang1Yu Huang2Yu Huang3Dan Gu4Dan Gu5Dan Gu6Dan Gu7Han Xue8Han Xue9Han Xue10Han Xue11Jinyan Yu12Jinyan Yu13Jinyan Yu14Jinyan Yu15Yuanyue Tang16Yuanyue Tang17Yuanyue Tang18Yuanyue Tang19Jinlin Huang20Jinlin Huang21Jinlin Huang22Jinlin Huang23Yunzeng Zhang24Yunzeng Zhang25Yunzeng Zhang26Yunzeng Zhang27Xinan Jiao28Xinan Jiao29Xinan Jiao30Xinan Jiao31Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, ChinaJiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, ChinaJoint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, ChinaKey Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, ChinaJiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, ChinaJoint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, ChinaKey Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, ChinaJiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, ChinaJoint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, ChinaKey Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, ChinaJiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, ChinaJoint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, ChinaKey Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, ChinaJiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, ChinaJoint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, ChinaKey Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, ChinaJiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, ChinaJoint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, ChinaKey Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, ChinaJiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, ChinaJoint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, ChinaKey Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, ChinaJiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou, ChinaJiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, ChinaJoint International Research Laboratory of Agriculture and Agri-product Safety of the Ministry of Education, Yangzhou University, Yangzhou, ChinaKey Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China, Yangzhou University, Yangzhou, ChinaCampylobacter jejuni is among the most prevalent foodborne zoonotic pathogens leading to diarrheal diseases. In this study, we developed a CRISPR-Cas12b-based system to rapidly and accurately detect C. jejuni contamination. Identification of C. jejuni-specific and -conserved genomic signatures is a fundamental step in development of the detection system. By comparing C. jejuni genome sequences with those of the closely related Campylobacter coli, followed by comprehensive online BLAST searches, a 20-bp C. jejuni-conserved (identical in 1024 out of 1037 analyzed C. jejuni genome sequences) and -specific (no identical sequence detected in non-C. jejuni strains) sequence was identified and the system was then assembled. In further experiments, strong green fluorescence was observed only when C. jejuni DNA was present in the system, highlighting the specificity of this system. The assay, with a sample-to-answer time of ∼40 min, positively detected chicken samples that were contaminated with a dose of approximately 10 CFU C. jejuni per gram of chicken, which was >10 times more sensitive than the traditional Campylobacter isolation method, suggesting that this method shows promise for onsite C. jejuni detection. This study provides an example of bioinformatics-guided CRISPR-Cas12b-based detection system development for rapid and accurate onsite pathogen detection.https://www.frontiersin.org/articles/10.3389/fmicb.2021.649010/fullCampylobacter jejuniCRISPR-Cas12bprotospacer discoverycomparative genomicsvisualized detection |
| spellingShingle | Yu Huang Yu Huang Yu Huang Yu Huang Dan Gu Dan Gu Dan Gu Dan Gu Han Xue Han Xue Han Xue Han Xue Jinyan Yu Jinyan Yu Jinyan Yu Jinyan Yu Yuanyue Tang Yuanyue Tang Yuanyue Tang Yuanyue Tang Jinlin Huang Jinlin Huang Jinlin Huang Jinlin Huang Yunzeng Zhang Yunzeng Zhang Yunzeng Zhang Yunzeng Zhang Xinan Jiao Xinan Jiao Xinan Jiao Xinan Jiao Rapid and Accurate Campylobacter jejuni Detection With CRISPR-Cas12b Based on Newly Identified Campylobacter jejuni-Specific and -Conserved Genomic Signatures Frontiers in Microbiology Campylobacter jejuni CRISPR-Cas12b protospacer discovery comparative genomics visualized detection |
| title | Rapid and Accurate Campylobacter jejuni Detection With CRISPR-Cas12b Based on Newly Identified Campylobacter jejuni-Specific and -Conserved Genomic Signatures |
| title_full | Rapid and Accurate Campylobacter jejuni Detection With CRISPR-Cas12b Based on Newly Identified Campylobacter jejuni-Specific and -Conserved Genomic Signatures |
| title_fullStr | Rapid and Accurate Campylobacter jejuni Detection With CRISPR-Cas12b Based on Newly Identified Campylobacter jejuni-Specific and -Conserved Genomic Signatures |
| title_full_unstemmed | Rapid and Accurate Campylobacter jejuni Detection With CRISPR-Cas12b Based on Newly Identified Campylobacter jejuni-Specific and -Conserved Genomic Signatures |
| title_short | Rapid and Accurate Campylobacter jejuni Detection With CRISPR-Cas12b Based on Newly Identified Campylobacter jejuni-Specific and -Conserved Genomic Signatures |
| title_sort | rapid and accurate campylobacter jejuni detection with crispr cas12b based on newly identified campylobacter jejuni specific and conserved genomic signatures |
| topic | Campylobacter jejuni CRISPR-Cas12b protospacer discovery comparative genomics visualized detection |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2021.649010/full |
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