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Open AccessArticle

Development of PCR, LAMP and qPCR Assays for the Detection of Aflatoxigenic Strains of Aspergillus flavus and A. parasiticus in Hazelnut

1
Centre of Competence for the Innovation in the Agro-Environmental Sector—AGROINNOVA, University of Turin, via Paolo Braccini 2, I-10095 Grugliasco, Italy
2
Department of Agricultural, Forestry and Food Sciences (DiSAFA), University of Torino, via Paolo Braccini 2, I-10095 Grugliasco, Italy
*
Author to whom correspondence should be addressed.
Toxins 2020, 12(12), 757; https://doi.org/10.3390/toxins12120757
Received: 30 September 2020 / Revised: 23 November 2020 / Accepted: 26 November 2020 / Published: 30 November 2020
(This article belongs to the Collection Aflatoxins)
Aspergillus flavus and A. parasiticus are two species able to produce aflatoxins in foodstuffs, and in particular in hazelnuts, at harvest and during postharvest phase. As not all the strains of these species are aflatoxin producers, it is necessary to develop techniques that can detect aflatoxigenic from not aflatoxigenic strains. Two assays, a LAMP (loop-mediated isothermal amplification) and a real time PCR with TaqMan® probe were designed and validated in terms of specificity, sensitivity, reproducibility, and repeatability. The capability of the strains to produce aflatoxins was measured in vitro and both assays showed to be specific for the aflatoxigenic strains of A. flavus and A. parasiticus. The limit of detection of the LAMP assay was 100–999 picograms of DNA, while the qPCR detected 160 femtograms of DNA in hazelnuts. Both techniques were validated using artificially inoculated hazelnuts and naturally infected hazelnuts. The qPCR was able to detect as few as eight cells of aflatoxigenic Aspergillus in naturally infected hazelnut. The combination of the LAMP assay, which can be performed in less than an hour, as screening method, with the high sensitivity of the qPCR, as confirmation assay, is able to detect aflatoxigenic strains already in field, helping to preserve the food safety of hazelnuts. View Full-Text
Keywords: LAMP; qPCR; detection; hazelnuts; Aspergillus flavus; A. parasiticus; aflatoxins LAMP; qPCR; detection; hazelnuts; Aspergillus flavus; A. parasiticus; aflatoxins
MDPI and ACS Style

Ortega, S.F.; Siciliano, I.; Prencipe, S.; Gullino, M.L.; Spadaro, D. Development of PCR, LAMP and qPCR Assays for the Detection of Aflatoxigenic Strains of Aspergillus flavus and A. parasiticus in Hazelnut. Toxins 2020, 12, 757.

AMA Style

Ortega SF, Siciliano I, Prencipe S, Gullino ML, Spadaro D. Development of PCR, LAMP and qPCR Assays for the Detection of Aflatoxigenic Strains of Aspergillus flavus and A. parasiticus in Hazelnut. Toxins. 2020; 12(12):757.

Chicago/Turabian Style

Ortega, Sara F.; Siciliano, Ilenia; Prencipe, Simona; Gullino, Maria L.; Spadaro, Davide. 2020. "Development of PCR, LAMP and qPCR Assays for the Detection of Aflatoxigenic Strains of Aspergillus flavus and A. parasiticus in Hazelnut" Toxins 12, no. 12: 757.

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