Development of real-time PCR methods to quantify patulin-producing molds in food products

Rodríguez, Alicia; Luque, M. Isabel; Andrade, María J.; Rodrı́guez, Mar; Asensio, Miguel A.; Córdoba, Juan J.

doi:10.1016/j.fm.2011.04.004

Cited by 46 publications

(19 citation statements)

References 34 publications

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“…The suitable values of PCR efficiency (104.72%,~100%, calculated by the equation E = 10 −1/slope − 1) and R 2 (0.9997) of standard curve were observed (Fig. 3A), indicating that the qPCR assay developed was accurate (Martinez-Blanch et al, 2009;Rodríguez et al, 2011). Fig.…”

Section: Population Of C Kluyveri In Pit Mudsmentioning

confidence: 80%

Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production

2015

International Journal of Food Microbiology

113

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Section: Population Of C Kluyveri In Pit Mudsmentioning

confidence: 80%

Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production

2015

International Journal of Food Microbiology

113

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“…The extractions of genomic DNA for qPCR analysis were performed from 20 g portions of the same samples as those used for PAT extraction and analysis. Extractions were performed according to the protocol described by Rodriguez et al (2011) with slight modifications. Fifty milliliters of Tris-HCl (pH 8) were added to 20 g of homogenized sample and then vortexed for homogenization.…”

Section: Methodsmentioning

confidence: 99%

Biocontrol Agents Increase the Specific Rate of Patulin Production by Penicillium expansum but Decrease the Disease and Total Patulin Contamination of Apples

et al. 2017

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Synthetic fungicides are commonly employed for the control of postharvest diseases of fruits. However, due to health concerns about the use of these chemicals, alternative control methods including biocontrol based on antagonistic yeasts are gaining in popularity. In this study, we investigated the effects of two biocontrol yeasts, Rhodotorula mucilaginosa strain 3617 and Rhodotorula kratochvilovae strain LS11, on blue mold and patulin (PAT) contamination caused by Penicillium expansum strains PY and FS7 in artificially inoculated Fuji apples stored at 20°C for 9 days. To correlate the development of the P. expansum strains in yeast-treated and untreated apples with PAT production, we quantified their biomass in the infected fruits using a recently published quantitative real-time polymerase chain reaction method based on specific primers for patF, a gene from P. expansum that is involved in PAT biosynthesis. Both yeasts significantly reduced the disease incidence caused by the two strains of P. expansum up to 5–7 days of incubation, and lowered their biomass and the progression of symptoms up to 9 days. Interestingly, both yeasts strains increased the rate of PAT production (expressed as ng patulin/μg fungal DNA) by the two pathogenic strains. Nevertheless, both biocontrol agents reduced the total PAT contamination, especially in the case of P. expansum strain FS7, the higher PAT producer of the two tested P. expansum strains. Comparing between the yeast strains, R. kratochvilovae LS11 was more effective than R. mucilaginosa 3617 for the control of P. expansum.

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“…is reported to be 31 fg [29]. Moreover, this sensitivity level is much better than levels reported in literature concerning Penicillium detection in food matrices [17,18,19]. Indeed, by the introduction of the nested approach, it was possible to enhance the performance of a traditional real-time PCR assay.…”

Section: Discussionmentioning

confidence: 99%

“…from apples, sweet cherries, and table grapes [9]. Finally, qPCR assays have been set up targeting patulin biosynthetic genes in terms of presence and expression [18,19]. However, most of the molecular assays targeting pathogens in biological matrices suffer from difficulties in extracting DNA of good quality and quantity, and from low pathogen representation.…”

Section: Introductionmentioning

confidence: 99%

A Rapid Assay to Detect Toxigenic Penicillium spp. Contamination in Wine and Musts

Sanzani

Miazzi

Rienzo

et al. 2016

Toxins

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Wine and fermenting musts are grape products widely consumed worldwide. Since the presence of mycotoxin-producing fungi may greatly compromise their quality characteristics and safety, there is an increasing need for relatively rapid “user friendly” quantitative assays to detect fungal contamination both in grapes delivered to wineries and in final products. Although other fungi are most frequently involved in grape deterioration, secondary infections by Penicillium spp. are quite common, especially in cool areas with high humidity and in wines obtained by partially dried grapes. In this work, a single-tube nested real-time PCR approach—successfully applied to hazelnut and peanut allergen detection—was tested for the first time to trace Penicillium spp. in musts and wines. The method consisted of two sets of primers specifically designed to target the β-tubulin gene, to be simultaneously applied with the aim of lowering the detection limit of conventional real-time PCR. The assay was able to detect up to 1 fg of Penicillium DNA. As confirmation, patulin content of representative samples was determined. Most of analyzed wines/musts returned contaminated results at >50 ppb and a 76% accordance with molecular assay was observed. Although further large-scale trials are needed, these results encourage the use of the newly developed method in the pre-screening of fresh and processed grapes for the presence of Penicillium DNA before the evaluation of related toxins.

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Development of real-time PCR methods to quantify patulin-producing molds in food products

Cited by 46 publications

References 34 publications

Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production

Identification and quantification of the caproic acid-producing bacterium Clostridium kluyveri in the fermentation of pit mud used for Chinese strong-aroma type liquor production

Biocontrol Agents Increase the Specific Rate of Patulin Production by Penicillium expansum but Decrease the Disease and Total Patulin Contamination of Apples

A Rapid Assay to Detect Toxigenic Penicillium spp. Contamination in Wine and Musts

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