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Rapid and sensitive detection of Karenia mikimotoi by loop-mediated isothermal amplification combined with a lateral flow dipstick

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Abstract  

Harmful algal blooms frequently occur in various coastal regions worldwide, deteriorating marine ecology and causing huge economic losses. Therefore, developing a potential method for rapid detection of harmful algae species is highly necessitated. In this study, a loop-mediated isothermal amplification (LAMP) method coupled with a lateral flow dipstick (LFD) was developed for detecting the harmful algae Karenia mikimotoi. In this method, the internal transcribed spacer (ITS) sequence of K. mikimotoi was used as the template, and the corresponding specific primers were designed by the online software PrimerExplorer V5. Biotin was labeled on the 5′ end of forward inner primer (FIP), and the LAMP reaction was performed under the determined optimal conditions of 63℃ and 60 min. The lowest concentration of K. mikimotoi DNA tested using LAMP was 3.3 × 10–1 pg/μL. Additionally, a 6-FAM-labeled probe was designed and displayed on the LFD after hybridization of the amplified product with the probe. The results demonstrated that LAMP-LFD could be a promising approach for detecting and monitoring harmful algae due to its high sensitivity and specificity.

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Data Availability

All data generated or analyzed during this study are included in this published article.

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Funding

This work was funded by the Special Foundation for National Science and Technology Basic Research Program of China (Grant No. 2018FY100207); the National Key Research and Development Program of China (Grant No. 2017YFC1404304); the Strategic Priority Research Program of the Chinese Academy of Sciences (Grant No. XDA23050302); the Sino-Australian Centre for Healthy Coasts (Grant No. 2016YFE0101500); Aoshan Science and Technology Innovation Project, Qingdao National Laboratory for Marine Science and Technology (Grant No. 2016ASKJ02-1); Foundation of Key Laboratory of Integrated Marine Monitoring; and Applied Technologies for Harmful Algal Blooms (Grant No. MATHAB201707).

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Authors and Affiliations

  1. Changjiang River Estuary Ecosystem Research Station, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao, 266071, Shandong, China

    Xiaotian Han, Tian Yan & Rencheng Yu

  2. Qingdao National Laboratory for Marine Science and Technology, Qingdao, 266237, China

    Xiaotian Han, Tian Yan & Rencheng Yu

  3. Center for Ocean Mega-Science, Chinese Academy of Sciences, Qingdao, 266100, China

    Xiaotian Han, Tian Yan & Rencheng Yu

  4. Guangzhou, China

    Ting Zhao

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  1. Xiaotian Han
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  2. Ting Zhao
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  3. Tian Yan
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  4. Rencheng Yu
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Contributions

Han Xiaotian: supervision, conceptualization, project administration, and data curation. Zhao Ting: writing—original draft, writing—review and editing, methodology, and investigation. Yan Tian: methodology and investigation. Yu Rencheng: methodology and investigation.

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Correspondence to Xiaotian Han.

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Xiaotian Han and Ting Zhao contributed equally to this work and should be considered co-first authors.

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Han, X., Zhao, T., Yan, T. et al. Rapid and sensitive detection of Karenia mikimotoi by loop-mediated isothermal amplification combined with a lateral flow dipstick. Environ Sci Pollut Res 29, 24696–24703 (2022). https://doi.org/10.1007/s11356-021-17536-w

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  • DOI: https://doi.org/10.1007/s11356-021-17536-w

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