A Rapid Assay to Detect Toxigenic Penicillium spp. Contamination in Wine and Musts
Abstract
:1. Introduction
2. Results
2.1. Set up of Experimental Design
2.2. Specificity and Sensitivity Assay
2.3. Penicillium Detection in Real Samples
2.4. Patulin Quantification in Real Samples
3. Discussion
4. Conclusions
5. Materials and Methods
5.1. Sample Collection
5.2. DNA Extraction
5.3. Penicillium Detection System
5.4. Nested One-Tube Real-Time PCR Assays
5.5. Specificity and Sensitivity Assay
5.6. Patulin Evaluation
5.6.1. Chemicals and Reagents
5.6.2. Extraction Procedure
5.6.3. Chromatographic and Mass Spectrometric Conditions
5.7. Statistical Analysis
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Isolate Code | Organism | Presence of Amplification Product |
---|---|---|
Pex6 | Penicillium expansum | + |
Pex29 | Penicillium chrysogenum | + |
Pex30 | Penicillium crustosum | + |
A64 | Alternaria alternata | - |
FV52 | Botrytis cinerea | - |
FV509 | Monilia laxa | - |
FV139 | Phellinus ignarius | - |
FV366 | Sclerotinia sclerotiorum | - |
FV155 | Cladosporium spp. | - |
FV406 | Aspergillus spp. | - |
FV150 | Rhizopus stolonifer | - |
FV126 | Fusarium spp. | - |
VV1 | Vitis vinifera | - |
Sample Code | Type | Storage | DNA Concentration (pg/μL) |
---|---|---|---|
C10 | White must | Cistern | - |
C11a | White wine | Cistern | 1.007 |
C11b | White must | Cistern | - |
C12a | Red must | Cistern | 0.010 |
C12b | White must | Cistern | - |
C13 | White must | Cistern | - |
C14 | White must | Cistern | - |
C15 | White must | Cistern | - |
C19 | White must | Cistern | - |
C21 | White must | Cistern | - |
C22 | White must | Cistern | - |
C23 | White must | Cistern | - |
C35 | White wine | Cistern | - |
C43 | White must | Cistern | - |
C47 | White must | Cistern | - |
C48 | White must | Cistern | - |
C52 | White must | Cistern | - |
C53 | Red must | Cistern | - |
C55 | Red must | Cistern | 0.016 |
C56 | Red must | Cistern | - |
C57 | Red must | Cistern | 0.003 |
C59 | Red must | Cistern | 0.006 |
SS5 | White must | Silos | 0.011 |
SS8 | Red must | Silos | 0.029 |
SS10 | Red wine | Silos | 1.0074 |
SS13 | Red wine | Silos | - |
SS14 | Red must | Silos | 0.010 |
SS15 | White must | Silos | 0.002 |
SS17 | Red wine | Silos | 0.002 |
SS19 | Red wine | Silos | - |
SS21 | White wine | Silos | 0.002 |
SS25 | Red must | Silos | 1.96 |
SS26 | Red must | Silos | 2.634 |
SS27 | Red must | Silos | - |
SS28 | Red must | Silos | - |
SS29 | Red must | Silos | 0.014 |
SS33 | White must | Silos | 0.003 |
SS34 | White must | Silos | 0.010 |
SS36 | White wine | Silos | - |
SS39 | White must | Silos | 0.056 |
SS42 | Red must | Silos | - |
SS44 | Red wine | Silos | - |
SS45 | White wine | Silos | 0.002 |
SS47 | Red wine | Silos | - |
SS48 | Red wine | Silos | - |
SS51 | Red must | Silos | 0.056 |
SS52 | Red must | Silos | - |
SS73 | White wine | Silos | - |
SS75 | Red wine | Silos | - |
SS77 | Red wine | Silos | - |
T2 | Red wine | Tank | 0.034 |
T7 | Red must | Tank | 0.183 |
T11 | Red must | Tank | - |
T13 | Red must | Tank | 0.034 |
T15 | Red must | Tank | - |
T17 | White wine | Tank | 0.006 |
T20 | Red wine | Tank | - |
T21 | Red wine | Tank | 0.011 |
T23 | Red wine | Tank | 0.010 |
T24 | White wine | Tank | - |
T25 | Red wine | Tank | - |
T26 | Red wine | Tank | - |
T27 | Red wine | Tank | - |
T28 | Red wine | Tank | 0.065 |
T31 | Red wine | Tank | 0.070 |
T32 | White wine | Tank | 0.042 |
T33 | Red wine | Tank | 0.010 |
T35 | Red wine | Tank | 0.309 |
T36 | Red wine | Tank | - |
T38 | Red wine | Tank | 0.029 |
T40 | White wine | Tank | 0.001 |
T41 | White wine | Tank | 0.023 |
T44 | Red wine | Tank | - |
T45 | Red wine | Tank | - |
T48 | White wine | Tank | 0.014 |
T49 | Red wine | Tank | - |
T51 | Red wine | Tank | - |
T52 | Red must | Tank | - |
T55 | Red must | Tank | 0.016 |
T58 | Red wine | Tank | 0.010 |
T70 | Red wine | Tank | 0.070 |
Sample Code | Type | Penicillium DNA Concentration (pg/μL) | Patulin Concentration (μg/L) |
---|---|---|---|
SS8 | Red must | 0.029 | 0 |
SS10 | Red wine | 1.0074 | 173 |
SS14 | Red must | 0.010 | 277 |
SS17 | Red wine | 0.002 | 397 |
SS21 | White wine | 0.002 | 27 |
SS29 | Red must | 0.014 | 154 |
SS33 | White must | 0.003 | 778 |
SS34 | White must | 0.010 | 0 |
SS39 | White must | 0.056 | 65 |
T13 | Red must | 0.034 | 60 |
T17 | White wine | 0.006 | 1911 |
T21 | Red wine | 0.011 | 712 |
T35 | Red wine | 0.309 | 82 |
T40 | White wine | 0.001 | 681 |
T41 | White wine | 0.023 | 0 |
T48 | White wine | 0.014 | 0 |
T58 | Red wine | 0.010 | 669 |
ID | Q1 Mass (Da) | Q3 Mass (Da) | Dwell (msec) | DP | EP | CE | CXP |
---|---|---|---|---|---|---|---|
Patulin 1 | 152.9 | 109.0 | 5.00 | −45.00 | −10.00 | −13.00 | −7.00 |
Patulin 2 | 152.9 | 80.9 | 5.00 | −45.00 | −10.00 | −15.00 | −7.00 |
Dinoseb 1 | 239.0 | 133.9 | 5.00 | −120.00 | −8.00 | −58.68 | −10.00 |
Dinoseb 2 | 239.0 | 163.1 | 5.00 | −120.00 | −8.00 | −41.78 | −10.00 |
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Sanzani, S.M.; Miazzi, M.M.; Di Rienzo, V.; Fanelli, V.; Gambacorta, G.; Taurino, M.R.; Montemurro, C. A Rapid Assay to Detect Toxigenic Penicillium spp. Contamination in Wine and Musts. Toxins 2016, 8, 235. https://doi.org/10.3390/toxins8080235
Sanzani SM, Miazzi MM, Di Rienzo V, Fanelli V, Gambacorta G, Taurino MR, Montemurro C. A Rapid Assay to Detect Toxigenic Penicillium spp. Contamination in Wine and Musts. Toxins. 2016; 8(8):235. https://doi.org/10.3390/toxins8080235
Chicago/Turabian StyleSanzani, Simona Marianna, Monica Marilena Miazzi, Valentina Di Rienzo, Valentina Fanelli, Giuseppe Gambacorta, Maria Rosaria Taurino, and Cinzia Montemurro. 2016. "A Rapid Assay to Detect Toxigenic Penicillium spp. Contamination in Wine and Musts" Toxins 8, no. 8: 235. https://doi.org/10.3390/toxins8080235