A visual on-site method for African swine fever virus detection in raw pig tissues |
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| Authors: | Xiaoli Zhao Xiudan Wang Xuejiao Wang Sen Liu Hao Jing HaiXia Wang Chao Shi Cuiping Ma |
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| Affiliation: | 1. Key Laboratory of Optic-electric Sensing and Analytical Chemistry for Life Science, MOE, Shandong Provincial Key Laboratory of Biochemical Engineering, College of Marine Science and Biological Engineering, College of Chemistry and Molecular Engineering, Qingdao University of Science and Technology, Qingdao, People's Republic of China;2. Navid Biotech Co., Ltd., Qingdao, People's Republic of China;3. Qingdao Nucleic Acid Rapid Testing International Science and Technology Cooperation Base, College of Life Sciences, Department of Pathogenic Biology, School of Basic Medicine, and Department of Clinical Laboratory, The Affiliated Hospital of Qingdao University, Qingdao University, Qingdao, People's Republic of China |
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| Abstract: | The spread of African swine fever virus (ASFV) leads to high mortality in domestic pigs, and has caused huge losses in the global pork industry. Therefore, the rapid detection of ASFV is essential for the prevention and control of the epidemic. Herein, a denaturation bubble-mediated strand exchange amplification (SEA) with colorimetric detection strategy for the rapid detection of ASFV was established and the results were visible with the naked eyes. The entire detection process based on isothermal amplification was performed on a portable metal bath only. And the whole process including rapid extraction and visual detection was finished within 50 min. The limit of detection of this method reached to 104 copies/μl of plasmid containing p72 gene. And the ASFV was successfully detected in eight different tissue and seven types of pig feed which revealed a strong anti-interference ability in the presence of complex matrix. Through comparing 94 field positive and negative ASFV samples, excellent performance of SEA assay with 92.86% sensitivity and 100% specificity relative to PCR was observed. The developed method could be used for on-site detection of ASFV with simplicity, rapidity, and visuality, especially suitable for pork industry food safety and ASFV control in resource-limited areas. |
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