Quantification of Aspergillus carbonarius in grapes using a real time PCR assay

Tryfinopoulou, Paschalitsa; Kizis, Dimosthenis; Nychas, George‐John E.; Panagou, Efstathios Ζ.

doi:10.1016/j.fm.2015.05.009

Cited by 11 publications

(5 citation statements)

References 28 publications

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“…The sterilized blueberry was inoculated using a modification of the procedure described by Tryfinopoulou et al . (). Instead of inoculating with conidia, pieces (2 mm × 2 mm) of growing mycelia from potato dextrose agar cultures of each heat‐resistant fungi were used to inoculate the blueberry, which were then incubated at 25°C for 1–3 weeks.…”

Section: Methodsmentioning

confidence: 97%

Design of a species-specific PCR method for the detection of the heat-resistant fungi Talaromyces macrosporus and Talaromyces trachyspermus

Yamashita

Nakagawa²,

Sakaguchi

et al. 2017

Lett Appl Microbiol

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Significance and Impact of the Study: Polymerase chain reaction (PCR)-based detection is rapid, convenient and sensitive compared with traditional methods of detecting heat-resistant fungi. In this study, a PCR-based method was developed for the detection and identification of amplification products from Talaromyces macrosporus and Talaromyces trachyspermus using primer sets that target the isocitrate lyase gene. This method could be used for the on-site detection of T. macrosporus and T. trachyspermus in the near future, and will be helpful in the safety control of raw materials and in food and beverage production. AbstractHeat-resistant fungi occur sporadically and are a continuing problem for the food and beverage industry. The genus Talaromyces, as a typical fungus, is capable of producing the heat-resistant ascospores responsible for the spoilage of processed food products. Isocitrate lyase, a signature enzyme of the glyoxylate cycle, is required for the metabolism of non-fermentable carbon compounds, like acetate and ethanol. Here, species-specific primer sets for detection and identification of DNA derived from Talaromyces macrosporus and Talaromyces trachyspermus were designed based on the nucleotide sequences of their isocitrate lyase genes. Polymerase chain reaction (PCR) using a species-specific primer set amplified products specific to T. macrosporus and T. trachyspermus. Other fungal species, such as Byssochlamys fulva and Hamigera striata, which cause food spoilage, were not detected using the Talaromyces-specific primer sets. The detection limit for each species-specific primer set was determined as being 50 pg of template DNA, without using a nested PCR method. The specificity of each species-specific primer set was maintained in the presence of 1,000-fold amounts of genomic DNA from other fungi. The method also detected fungal DNA extracted from blueberry inoculated with T. macrosporus. This PCR method provides a quick, simple, powerful and reliable way to detect T. macrosporus and T. trachyspermus.

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Section: Methodsmentioning

confidence: 97%

Design of a species-specific PCR method for the detection of the heat-resistant fungi Talaromyces macrosporus and Talaromyces trachyspermus

Yamashita

Nakagawa²,

Sakaguchi

et al. 2017

Lett Appl Microbiol

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“…A. carbonarius and members of the A. niger aggregate are main OTA producers, with the former being the predominant species responsible for OTA contamination in grapes and wine, due to the ability of almost all its strains to produce high levels of the toxin [179][180][181]. Several Aspergilli, including A. flavus, A. parasiticus, and A. nominus, are potent producers of aflatoxins in different food commodities, including grapes [182]. Fumonisins (mainly FB 2 ) are also produced by Aspergillus niger strains and occur naturally in grape and must [183,184].…”

Section: Molecular Methods For the Detection Of Mycotoxigenic Fungimentioning

confidence: 99%

Recent Advances in Mycotoxin Analysis and Detection of Mycotoxigenic Fungi in Grapes and Derived Products

Kizis

Vichou

Natskoulis³

2021

Sustainability

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Mycotoxins are secondary metabolites of filamentous fungi that can cause toxic effects in human and animal health. Most of the filamentous fungi that produce these mycotoxins belong to four genera, namely, Aspergillus, Penicillium, Fusarium, and Alternaria. Mycotoxigenic fungi, along with mycotoxins, create a constant and serious economic threat for agriculture in many terms, counting product losses due to crop contamination and food spoilage, as well malnutrition when considering nutritional quality degradation. Given the importance of robust and precise diagnostics of mycotoxins and the related producing fungi in the grape food chain, one of the most important agricultural sectors worldwide, the present review initially delivers a comprehensive presentation of mycotoxin reports on grape and derived products, including a wide range of commodities such as fresh grapes, raisins, wine, juices, and other processed products. Next, based on worldwide regulations’ requirements for mycotoxins, and referring to the relative literature, this work presents methodological approaches for mycotoxin determination, and stresses major methods for the detection of fungal species responsible for mycotoxin production. The principle of function and basic technical background on the available analytical and molecular biology techniques developed—including chromatography, mass spectrometry, immunochemical-based assays, biosensors, and molecular assays—is briefly given, and references for their application to grape and derived product testing are highlighted.

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“…DNA extractions from lyophilized mycelia were performed using the NucleoSpin ® plant DNA kit (Macherey‐Nagel, Duren, Germany) according to the manufacturer's instructions. Total genomic DNA extraction from grapes inoculated with conidia was performed using the Nucleospin ® food (Macherey‐Nagel) DNA extraction kit, according to the manufacturer's instructions, with modifications …”

Section: Methodsmentioning

confidence: 99%

“…The need for differentiation and taxonomic classification of known and new or cryptic species among black aspergilli, the necessity for rapid and accurate determination of certain Aspergillus species belonging to section Nigri – some of which have particular economic and agronomic interest – has led to the development of numerous polymerase chain reaction (PCR) methods. A considerable number of endpoint and real‐time PCR assays, either simplex or multiplex, have been developed during the last two decades, in order to detect and quantify these species in different food commodities and particularly in table and wine grapes . Within this scope, an early determination of the most common and principal OTA producing black aspergilli – such as A. carbonarius and other members of the A. niger aggregate – in grapes is a prerequisite for controlling product quality and toxin levels in grapes and grape products.…”

Section: Introductionmentioning

confidence: 99%

“…A considerable number of endpoint and real-time PCR assays, either simplex or multiplex, have been developed during the last two decades, in order to detect and quantify these species in different food commodities and particularly in table and wine grapes. [15][16][17][18][19][20][21][22][23][24][25][26] Within this scope, an early determination of the most common and principal OTA producing black aspergilli -such as A. carbonarius and other members of the A. niger aggregate -in grapes is a prerequisite for controlling product quality and toxin levels in grapes and grape products.…”

Section: Introductionmentioning

confidence: 99%

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Rapid and accurate identification of black aspergilli from grapes using high‐resolution melting (HRM) analysis

et al. 2018

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Quantification of Aspergillus carbonarius in grapes using a real time PCR assay

Cited by 11 publications

References 28 publications

Design of a species-specific PCR method for the detection of the heat-resistant fungi Talaromyces macrosporus and Talaromyces trachyspermus

Design of a species-specific PCR method for the detection of the heat-resistant fungi Talaromyces macrosporus and Talaromyces trachyspermus

Recent Advances in Mycotoxin Analysis and Detection of Mycotoxigenic Fungi in Grapes and Derived Products

Rapid and accurate identification of black aspergilli from grapes using high‐resolution melting (HRM) analysis

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