A Rapid and Efficient Loop-mediated Isothermal Amplification (LAMP) Assay for the Authentication of Food Supplements Based on Maitake (Grifola Frondosa)

Abstract

Grifola frondosa (“Maitake”) is an edible fungus with several nutraceutical properties, largely used in traditional medicine. The increased use of Maitake as a food supplements ingredient raised the need of accurate authentication methods since the morphological identification of G. frondosa is not feasible in formulated food supplements. We developed a diagnostic tool based on loop-mediated isothermal AMPlification (LAMP) for the detection of G. frondosa in food supplements. First, a modified CTAB protocol for DNA extraction from food supplements has been set up and it has been shown to be able to isolate amplifiable total genomic material from different types of commercial products. Subsequently, the LAMP assay confirmed high specificity and good analytical sensitivity, allowing to detect up to 0.62 pg of genomic DNA in less than 20 min. Ten related fungal species resulted negative, confirming the specificity of the assay. The presence of Maitake in commercial food supplements was confirmed, except for one, revealing a mislabeling (or a food fraud). This assay proved to be a rapid powerful tool for food authentication purposes and routine inspections at any level of the supply chain of Maitake-based products and it can be used as a model for other quality control assays of fungal food products. Supplementary Information The online version contains supplementary material available at 10.1007/s12161-022-02235-0.

Field ready: Development of a rapid LAMP‐based colorimetric assay for the causal agent of white pine blister rust, Cronartium ribicola

“…These results suggest that the putative matrix effect of components present in bark extract does not decrease the sensitivity of the LAMP assay at 40 pg or higher of DNA. Other studies have shown sufficient tolerance of LAMP assays to potential inhibitors present in host extract (Fochi et al, 2022; Kaneko et al, 2007), making it an efficient detection method without the need for lab‐based DNA purification.…”

“…Other studies have shown sufficient tolerance of LAMP assays to potential inhibitors present in host extract (Fochi et al, 2022;Kaneko et al, 2007), making it an efficient detection method without the need for lab-based DNA purification.…”

Horizon scan of DNA-based methods for quality control and monitoring of herbal preparations

“…( Sasaki and Nagumo, 2007 ; Chaudhary et al, 2012 ; Li et al, 2013 ; Chaudhary and Khan, 2014 ; Lai et al, 2015 ). While Zhao et al (2016) developed ITS2-specific LAMP primers targeting Crocus sativus L. (saffron) to enable simple and sensitive differentiation from adulterants, Fochi et al (2022) developed a LAMP-based diagnostic protocol for the authentication of food supplements made from the edible fungus Grifola frondosa (Dicks.) Gray (“Maitake”).…”