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Detection of microcystin producing cyanobacteria in Spirulina dietary supplements using multiplex HRM quantitative PCR

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Abstract

Arthrospira species, under the name ‘Spirulina’, are used as food supplement for its protein, vitamins, and minerals which have several health benefits. Cyanobacterial toxins including microcystins can possibly contaminate these dietary supplements causing hepatotoxicity, tumour formation, and other disorders. The safe use of dietary supplements necessitates the need to assess such toxins in the algal food supplement. The methods which evaluate these dietary supplements should be highly sensitive, cost-effective, and rapid. In this study, multiplex HRM qPCR analysis was used to detect microcystin (MC)-producing cyanobacteria in Spirulina dietary supplements. The multiplex HRM qPCR detection limit was found to be 25 ag of mcyB spiked in a standard concentration of pcb (25 pg). Two distinct melt curves characteristic of pcb (Tm 82.8 ± 0.07 °C) and mcyB (Tm 77.9 ± 0.05 °C) were observed. Microcystin contamination was detected only in the fish food supplements and not in human dietary supplements of Spirulina. Liquid chromatography–high-resolution mass spectrometry analysis further confirmed the presence of the congeners of microcystin in the identified positive samples.

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Acknowledgments

We would like to acknowledge the support and facility given by SRM University in carrying out this project.

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Authors and Affiliations

  1. Department of Genetic Engineering, SRM University, Kattankulathur, 603203, Kancheepuram District, Tamil Nadu, India

    Kamath Mukund Manali, Rex Arunraj & Mohandass Ramya

  2. Analytical Research and Development Division, Piramal Enterprises Limited, Ennore, Chennai, Tamil Nadu, India

    Thangarathinam Kumar

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  1. Kamath Mukund Manali
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  2. Rex Arunraj
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  3. Thangarathinam Kumar
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  4. Mohandass Ramya
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Correspondence to Mohandass Ramya.

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Manali, K.M., Arunraj, R., Kumar, T. et al. Detection of microcystin producing cyanobacteria in Spirulina dietary supplements using multiplex HRM quantitative PCR. J Appl Phycol 29, 1279–1286 (2017). https://doi.org/10.1007/s10811-016-1011-4

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