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Recombinase polymerase amplification combined with lateral flow dipstick for the rapid detection of Chattonella marina

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Abstract

Chattonella marina (Raphidophyceae) is a common microalgal species that has frequently caused harmful algal blooms (HABs). Here, recombinase polymerase amplification (RPA) was combined with a lateral flow dipstick (RPA-LFD) to establish a rapid detection method for this alga, aimed at providing an effective early warning of C. marina-forming HABs. A pair of specific RPA primers and an LFD probe was designed to target the internal transcribed spacer (ITS) sequence of C. marina. The specificity test with 20 control microalgal species indicated that the RPA-LFD was highly specific. Sensitivity test showed that the established RPA-LFD was 10 − 100 times more sensitive than conventional PCR. The detection limit of RPA-LFD was as low as 9.5 × 10−1 ng µL−1 for genomic DNA, 1.17 × 103 copies μL−1 for recombinant plasmid containing the inserted ITS sequence, and 10 cells mL−1 for the crude DNA extract of C. marina. In addition, the RPA-LFD was proved to be more accurate and sensitive than conventional PCR in the detection of target cells from natural samples, exhibiting its ability to be applied in the actual in situ detection scenarios. In conclusion, the developed RPA-LFD is rapid, specific, sensitive, and promising for being used in the point-of-care testing.

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Acknowledgements

This work was supported by Shandong Provincial Natural Science Foundation, China (ZR2020MD081); the National Scientific Foundation of China (No. 31600309, 41476086); HIT Scientific Research Innovation Fund/the Fundamental Research Funds for the Central Universities (No. HIT.NSRIF.201702 and HIT.NSRIF.201709); and HIT Environment and Ecology Innovation Special Funds (No. HSCJ201622).

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Authors and Affiliations

  1. School of Marine Science and Technology, Harbin Institute of Technology (Weihai), Wenhua West Road, 2#, Weihai, 264209, Shandong Province, People’s Republic of China

    Chunyun Zhang, Yuchen Yang, Fuguo Liu, Yuanyuan Wang & Guofu Chen

  2. School of Environment, Harbin Institute of Technology, Harbin, 264209, People’s Republic of China

    Fuguo Liu & Guofu Chen

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  1. Chunyun Zhang
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  2. Yuchen Yang
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  3. Fuguo Liu
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Zhang, C., Yang, Y., Liu, F. et al. Recombinase polymerase amplification combined with lateral flow dipstick for the rapid detection of Chattonella marina. J Appl Phycol 34, 1607–1620 (2022). https://doi.org/10.1007/s10811-022-02737-8

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