Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis
Porcine epidemic diarrhoea virus (PEDV) is a member of the genus Alphacoronavirus in the family Coronaviridae. It causes acute watery diarrhoea and vomiting in piglets with high a mortality rate. Currently, the GII genotype, PEDV, possesses a high separation rate in wild strains and is usually repor...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-06-01
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| Series: | Frontiers in Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fmicb.2022.920801/full |
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| author | Bingxu Qian Bingxu Qian Kai Liao Dexin Zeng Dexin Zeng Wanqing Peng Xiaodong Wu Jinming Li Zongyi Bo Yongxin Hu Wenlong Nan Yuan Wen Yuying Cao Feng Xue Xiaorong Zhang Jianjun Dai Jianjun Dai |
| author_facet | Bingxu Qian Bingxu Qian Kai Liao Dexin Zeng Dexin Zeng Wanqing Peng Xiaodong Wu Jinming Li Zongyi Bo Yongxin Hu Wenlong Nan Yuan Wen Yuying Cao Feng Xue Xiaorong Zhang Jianjun Dai Jianjun Dai |
| author_sort | Bingxu Qian |
| collection | DOAJ |
| description | Porcine epidemic diarrhoea virus (PEDV) is a member of the genus Alphacoronavirus in the family Coronaviridae. It causes acute watery diarrhoea and vomiting in piglets with high a mortality rate. Currently, the GII genotype, PEDV, possesses a high separation rate in wild strains and is usually reported in immunity failure cases, which indicates a need for a portable and sensitive detection method. Here, reverse transcription–recombinase aided amplification (RT-RAA) was combined with the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas12a system to establish a multiplexable, rapid and portable detection platform for PEDV. The CRISPR RNA (crRNA) against Spike (S) gene of GII PEDV specifically were added into the protocol. This system is suitable for different experimental conditions, including ultra-sensitive fluorescence, visual, UV light, or flow strip detection. Moreover, it exhibits high sensitivity and specificity and can detect at least 100 copies of the target gene in each reaction. The CRISPR/Cas12a detection platform requires less time and represents a rapid, reliable and practical tool for the rapid diagnosis of GII genotype PEDV. |
| first_indexed | 2024-12-11T16:17:45Z |
| format | Article |
| id | doaj.art-00109cd590bd4b3aaa52efdcc0a617a0 |
| institution | Directory Open Access Journal |
| issn | 1664-302X |
| language | English |
| last_indexed | 2024-12-11T16:17:45Z |
| publishDate | 2022-06-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Microbiology |
| spelling | doaj.art-00109cd590bd4b3aaa52efdcc0a617a02022-12-22T00:58:55ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2022-06-011310.3389/fmicb.2022.920801920801Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid DiagnosisBingxu Qian0Bingxu Qian1Kai Liao2Dexin Zeng3Dexin Zeng4Wanqing Peng5Xiaodong Wu6Jinming Li7Zongyi Bo8Yongxin Hu9Wenlong Nan10Yuan Wen11Yuying Cao12Feng Xue13Xiaorong Zhang14Jianjun Dai15Jianjun Dai16MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, ChinaNational Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, ChinaTechnology Center of Hefei Customs, Hefei, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, ChinaNational Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao, ChinaNational Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao, ChinaJoint International Research Laboratory of Agriculture and Agri-Product Safety, The Ministry of Education of China, Yangzhou University, Yangzhou, ChinaNational Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao, ChinaNational Research Center for Exotic Animal Diseases, China Animal Health and Epidemiology Center, Qingdao, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, ChinaJiangsu Co-Innovation Center for the Prevention and Control of Animal Infectious Disease and Zoonoses, College of Veterinary Medicine, Yangzhou University, Yangzhou, ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, ChinaSchool of Life Science and Technology, China Pharmaceutical University, Nanjing, ChinaPorcine epidemic diarrhoea virus (PEDV) is a member of the genus Alphacoronavirus in the family Coronaviridae. It causes acute watery diarrhoea and vomiting in piglets with high a mortality rate. Currently, the GII genotype, PEDV, possesses a high separation rate in wild strains and is usually reported in immunity failure cases, which indicates a need for a portable and sensitive detection method. Here, reverse transcription–recombinase aided amplification (RT-RAA) was combined with the Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)/Cas12a system to establish a multiplexable, rapid and portable detection platform for PEDV. The CRISPR RNA (crRNA) against Spike (S) gene of GII PEDV specifically were added into the protocol. This system is suitable for different experimental conditions, including ultra-sensitive fluorescence, visual, UV light, or flow strip detection. Moreover, it exhibits high sensitivity and specificity and can detect at least 100 copies of the target gene in each reaction. The CRISPR/Cas12a detection platform requires less time and represents a rapid, reliable and practical tool for the rapid diagnosis of GII genotype PEDV.https://www.frontiersin.org/articles/10.3389/fmicb.2022.920801/fullporcine epidemic diarrhoea virusGII genotypeCRISPR/Cas12arapid diagnosislateral flow strip |
| spellingShingle | Bingxu Qian Bingxu Qian Kai Liao Dexin Zeng Dexin Zeng Wanqing Peng Xiaodong Wu Jinming Li Zongyi Bo Yongxin Hu Wenlong Nan Yuan Wen Yuying Cao Feng Xue Xiaorong Zhang Jianjun Dai Jianjun Dai Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis Frontiers in Microbiology porcine epidemic diarrhoea virus GII genotype CRISPR/Cas12a rapid diagnosis lateral flow strip |
| title | Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis |
| title_full | Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis |
| title_fullStr | Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis |
| title_full_unstemmed | Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis |
| title_short | Clustered Regularly Interspaced Short Palindromic Repeat/Cas12a Mediated Multiplexable and Portable Detection Platform for GII Genotype Porcine Epidemic Diarrhoea Virus Rapid Diagnosis |
| title_sort | clustered regularly interspaced short palindromic repeat cas12a mediated multiplexable and portable detection platform for gii genotype porcine epidemic diarrhoea virus rapid diagnosis |
| topic | porcine epidemic diarrhoea virus GII genotype CRISPR/Cas12a rapid diagnosis lateral flow strip |
| url | https://www.frontiersin.org/articles/10.3389/fmicb.2022.920801/full |
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