Development of sensitive and specific multiplex PCR method for the detection of microcystin producing cyanobacteria in spirulina food supplements

Manali, Kamath Mukund; Arunraj, Rex; Ramakrishnan, Gautham Subramaniam; Ramya, Mohandass

doi:10.1007/s10068-018-0476-0

Cited by 10 publications

(8 citation statements)

References 34 publications

(34 reference statements)

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“…For one, the concentration determined fluctuated around the regulatory limit of 1000 µg/kg as stipulated in the Belgian Royal Decree 7 , 8 . The presence of microcystin toxins in algae FS has been described previously 3 , 18 , 20 .…”

Section: Resultsmentioning

confidence: 99%

Multi-Class UHPLC-MS/MS Method for Plant Toxins and Cyanotoxins in Food Supplements and Application for Belgian Market Samples

et al. 2021

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The presence of plant toxins and/or cyanotoxins in food supplements implies consumer health risks. Therefore, a targeted ultra-high performance liquid chromatographic-tandem mass spectrometric method to detect/quantify 25 toxins simultaneously in food supplement formulations was developed and validated. Full validation for tablets/powders and secondary validation for a liquid and soft gel capsule indicated that most compounds were efficiently extracted (≥ 75%), while others were only partly extracted (18 – 61%). Trueness was fulfilled (70 – 120%), with some exceptions (mostly at the lowest validation level). Intralaboratory repeatability, intra- and interlaboratory reproducibility values of ≤ 20%, ≤ 25%, and ≤ 25% were obtained for most, respectively. Matrix effects were found to be significant for most compounds. Good sensitivity (µg/kg level) was observed for galegin(e), lycopsamine, lycorine, rubiadin, skimmiamine, and vascin(e), in contrast to helveticoside, lucidin, lucidin-3-primveroside, plumbagin(e), and thujone, which were detected at the mg/kg level. The other compounds were characterized by a sensitivity between 10 to 1000 µg/kg. The validated methodology was applied for 52 food supplements (tablets, capsules, liquids/syrup, etc.) purchased from the Belgian market. In more than 25% of the samples, one or more toxins were detected (concentrations determined using standard addition). Lycopsamine, microcystin LR, solamargine, thujone, and vasicin(e) were the most frequently detected toxins. A clear link between the toxins detected and the plant species on the food supplement ingredient list could not always be established. This generic “dilute-and-shoot” procedure can be used for further research on toxins in food supplements and by extension other plant/algae-based food/feed commodities (herbs, edible flowers, etc.).

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Section: Resultsmentioning

confidence: 99%

Multi-Class UHPLC-MS/MS Method for Plant Toxins and Cyanotoxins in Food Supplements and Application for Belgian Market Samples

et al. 2021

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“…In addition, the microcystin toxin can accumulate in biomass under improper cultivation conditions. Therefore strict quality control is required to monitor food supplements with spirulina [4].…”

Section: The Use Of Spirulina In the Food Industrymentioning

confidence: 99%

“…Chlorella, Dunaliella salina, and Spirulina are the most significant interest in food production. At the same time, Spirulina platensis occupies a special place among industrial-scale microalgae in terms of the volume of products produced from it [4].…”

Section: Introductionmentioning

confidence: 99%

Processing microalgae for use as a suppliment in food industry

Iskusnykh

2022

IOP Conf. Ser.: Earth Environ. Sci.

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Raw bioresources for food production are the object of attention and study worldwide. Protein deficiency in nutrition is a global problem that is difficult to solve based on traditional crop and livestock resources. Microalgae is a valuable protein-mineral-vitamin supplement to human food, capable of increasing the adaptive potential of the body. The advantage of microalgae as a source of protein and essential micronutrients lies in the economy of their cultivation using simple nutrient media. Microalgae are promising for use as functional food products both in biomass (powders, syrups) and as sources of biologically active substances. Unlocking the potential of microalgae requires advanced methods of biotechnology and synthetic biology, which is associated with the need to isolate and modify the secondary metabolites of cyanobacteria to obtain a wide range of biologically active ingredients. Microalgae is a promising dietary supplement for creating innovative food products that are beneficial to health. The development of innovative food products containing microalgae requires the complex work of scientists and practitioners from all branches of the food industry. We are currently beginning a journey towards creating products enriched with microalgae.

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“…One is to detect MCs, including high-performance liquid chromatography (HPLC) (Hemmati et al 2019), liquid chromatography-mass spectrometry (LC-MS) (Palagama et al 2018), high-performance capillary electrophoresis (Massey et al 2020), enzyme-linked immunosorbent assay (ELISA) (Lu et al 2020a(Lu et al , 2020b, protein phosphatase inhibition assay, and biosensors (Moore et al 2016). The other is to detect MC synthesis genes (e.g., mcyA, mcyB, mcyE, and mcyG), such as polymerase chain reaction (PCR) (Manali et al 2019), quantitative PCR (qPCR; Otten et al 2017), and loop-mediated isothermal amplification (LAMP) (Ramya et al 2018). These methods can meet the need for highly sensitive detection of MCs producing cyanobacteria and have important reference significance for the detection of the MC synthesis genes.…”

mentioning

confidence: 99%

Rapid detection ofmcyGgene of microcystins producing cyanobacteria in water samples by recombinase polymerase amplification combined with lateral flow strips

Feng

2021

Journal of Water and Health

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Nowadays, cyanobacteria blooms and microcystins (MCs) pollution are threatening water safety and public health. In this study, a rapid detection method was established for detecting MCs producing cyanobacteria. The MC synthesis gene mcyG was measured through recombinase polymerase amplification combined with lateral flow strips (LF-RPA) technology. The target gene mcyG was amplified at a temperature range of 37–45 °C, and the amplification time to detect mcyG was only 15 min at 37 °C. The optimal reaction conditions were confirmed using single dependent variable experiments, suggesting that the best probe dosage for 50 μL of the reaction mixture was 0.2 μL, the best dilution ratio of products was 1/100, and the best loading volume was 10 μL. The specificity test proved that the LF-RPA assay could distinguish MCs producing cyanobacteria from nontoxic algae well. Within 35 min of amplification time, the detection limit of the LF-RPA assay was 103 copies/mL mcyG and 104 cells/mL Microcystis aeruginosa FACHB-905. Overall, the LF-RPA assay could detect MCs producing cyanobacteria in water samples quickly and accurately, and it has a great promise to be applied for monitoring the MCs producing cyanobacteria blooms in natural waters.

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Development of sensitive and specific multiplex PCR method for the detection of microcystin producing cyanobacteria in spirulina food supplements

Cited by 10 publications

References 34 publications

Multi-Class UHPLC-MS/MS Method for Plant Toxins and Cyanotoxins in Food Supplements and Application for Belgian Market Samples

Multi-Class UHPLC-MS/MS Method for Plant Toxins and Cyanotoxins in Food Supplements and Application for Belgian Market Samples

Processing microalgae for use as a suppliment in food industry

Rapid detection ofmcyGgene of microcystins producing cyanobacteria in water samples by recombinase polymerase amplification combined with lateral flow strips

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