| Autor: |
Chao Qin, Jiajia Liu, Wenqi Zhu, Muchu Zeng, Ke Xu, Jinmei Ding, Hao Zhou, Jianshen Zhu, Yuqing Ke, Lai Yan Li, Gaoyuan Sheng, Zhuoru Li, Huaixi Luo, Shengyao Jiang, Kangchun Chen, Xianting Ding, He Meng |
| Jazyk: |
angličtina |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
Frontiers in Veterinary Science, Vol 9 (2022) |
| Druh dokumentu: |
article |
| ISSN: |
2297-1769 |
| DOI: |
10.3389/fvets.2022.962438 |
| Popis: |
African swine fever virus (ASFV) is a leading cause of worldwide agricultural loss. ASFV is a highly contagious and lethal disease for both domestic and wild pigs, which has brought enormous economic losses to a number of countries. Conventional methods, such as general polymerase chain reaction and isothermal amplification, are time-consuming, instrument-dependent, and unsatisfactorily accurate. Therefore, rapid, sensitive, and field-deployable detection of ASFV is important for disease surveillance and control. Herein, we created a one-pot visual detection system for ASFV with CRISPR/Cas12a technology combined with LAMP or RPA. A mineral oil sealing strategy was adopted to mitigate sample cross-contamination between parallel vials during high-throughput testing. Furthermore, the blue fluorescence signal produced by ssDNA reporter could be observed by the naked eye without any dedicated instrument. For CRISPR-RPA system, detection could be completed within 40 min with advantageous sensitivity. While CRISPR-LAMP system could complete it within 60 min with a high sensitivity of 5.8 × 102 copies/μl. Furthermore, we verified such detection platforms display no cross-reactivity with other porcine DNA or RNA viruses. Both CRISPR-RPA and CRISPR-LAMP systems permit highly rapid, sensitive, specific, and low-cost Cas12a-mediated visual diagnostic of ASFV for point-of-care testing (POCT) applications. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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