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| Rapid identification of Pheretima aspergillum in earthworms using loop-mediated isothermal amplification technology |
| Hits 977 Download times 603 Received:January 10, 2021 |
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| DOI
10.11656/j.issn.1672-1519.2021.04.26 |
| Key Words
Pheretima aspergillum;loop-mediated isothermal amplification;molecular identification;rapid authentication |
| Author Name | Affiliation | E-mail | | ZHANG Huanyu | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | TAN Wei | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | GAO Han | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | WANG Wenxiu | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | YU Xiaolei | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | | | LI Zhenguo | Mudanjiang YouBo Pharmaceutical Co., Ltd., Mudanjiang 157000, China | lzgjzt@vip.163.com | | TIAN Xiaoxuan | Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China | tian_xiaoxuan@tjutcm.edu.cn |
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| Abstract
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| [Objective] Establishing the technology of loop-mediated isothermal amplification (LAMP) to realize the rapid detection of Pheretima aspergillum in earthworms.[Methods] We collected earthworms in different areas and extracted DNA of these specimens.Subsequently,PCR amplification,sequencing and alignment were performed based on the mitochondrial cytochrome c oxidase subunit I (COI) genes to identify thespecies of samples. We performed LAMP experiment using the newly designed primers. After reaction,the SYBR Green I dye was added in amplification product,and the specificity of primers were detected by observing the color changes under natural and ultraviolet light respectively,or analyzed by agarose gel electrophoresis.[Results] Pheretima aspergillum were successfully specific amplified using the newly designed LAMP primer setsand the detection limitup to 40 fg/μL,with highly sensitivity.[Conclusion] LAMP technology can be utilized to realize the field rapid detection of Pheretima aspergillum. |
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