Real-time PCR for quantification of toxigenic Fusarium species in cereals

Tuija Sarlin, Teija Koivula, M. Jestoi, A. Rizzo, S. Paavanen-Huhtala, T. Yli-Mattila, Auli Haikara

Research output: Contribution to conferenceOther conference contributionScientific

Abstract

Fusarium species are potential mycotoxin producers in cereals. In near future EU is going to set the maximum limit values for Fusarium toxins in raw cereals and cereal products. Mycotoxin analyses are expensive and time-consuming hence a rapid and reliable quantification method for toxigenic Fusarium species is needed in order to be able to evaluate the mycotoxin risk in cereal-based industry. We have applied real-time PCR technique for the quantification of trichothecene-producing Fusarium species present in barley and malt samples. PCR results were compared to the amount of trichothecenes determined in the samples. Furthermore, highly toxigenic Fusarium graminearum was quantified in cereals by real-time PCR. DNA was extracted from ground kernels using FastDNA Spin Kit for Soil and analysed in a LightCycler„¥ system using the double-stranded DNA ¡Vbinding fluorescent dye SYBR Green I or the fluorigenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. PCR results correlated with the trichothecene content of the grain samples. Non-specific products such as primer dimers were amplified in the SYBR Green I assay, which interfered the quantification. Non-specific amplicons were not detected with the TaqMan probes. In addition, this assay was regarded as sensitive and reproducible.
Original languageEnglish
Publication statusPublished - 2004
MoE publication typeNot Eligible
EventFOSARE Seminar Series 5 Contaminants and Influence of Agricultural Practices - Brussels, Belgium
Duration: 18 Mar 200419 Mar 2004

Seminar

SeminarFOSARE Seminar Series 5 Contaminants and Influence of Agricultural Practices
CountryBelgium
CityBrussels
Period18/03/0419/03/04

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Fusarium
quantitative polymerase chain reaction
trichothecenes
mycotoxins
grain products
fluorescent dyes
Fusarium graminearum
malt
DNA
assays
sampling
toxins
barley
industry
seeds
methodology
soil

Cite this

Sarlin, T., Koivula, T., Jestoi, M., Rizzo, A., Paavanen-Huhtala, S., Yli-Mattila, T., & Haikara, A. (2004). Real-time PCR for quantification of toxigenic Fusarium species in cereals. FOSARE Seminar Series 5 Contaminants and Influence of Agricultural Practices, Brussels, Belgium.
Sarlin, Tuija ; Koivula, Teija ; Jestoi, M. ; Rizzo, A. ; Paavanen-Huhtala, S. ; Yli-Mattila, T. ; Haikara, Auli. / Real-time PCR for quantification of toxigenic Fusarium species in cereals. FOSARE Seminar Series 5 Contaminants and Influence of Agricultural Practices, Brussels, Belgium.
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abstract = "Fusarium species are potential mycotoxin producers in cereals. In near future EU is going to set the maximum limit values for Fusarium toxins in raw cereals and cereal products. Mycotoxin analyses are expensive and time-consuming hence a rapid and reliable quantification method for toxigenic Fusarium species is needed in order to be able to evaluate the mycotoxin risk in cereal-based industry. We have applied real-time PCR technique for the quantification of trichothecene-producing Fusarium species present in barley and malt samples. PCR results were compared to the amount of trichothecenes determined in the samples. Furthermore, highly toxigenic Fusarium graminearum was quantified in cereals by real-time PCR. DNA was extracted from ground kernels using FastDNA Spin Kit for Soil and analysed in a LightCycler„¥ system using the double-stranded DNA ¡Vbinding fluorescent dye SYBR Green I or the fluorigenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. PCR results correlated with the trichothecene content of the grain samples. Non-specific products such as primer dimers were amplified in the SYBR Green I assay, which interfered the quantification. Non-specific amplicons were not detected with the TaqMan probes. In addition, this assay was regarded as sensitive and reproducible.",
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Sarlin, T, Koivula, T, Jestoi, M, Rizzo, A, Paavanen-Huhtala, S, Yli-Mattila, T & Haikara, A 2004, 'Real-time PCR for quantification of toxigenic Fusarium species in cereals' FOSARE Seminar Series 5 Contaminants and Influence of Agricultural Practices, Brussels, Belgium, 18/03/04 - 19/03/04, .

Real-time PCR for quantification of toxigenic Fusarium species in cereals. / Sarlin, Tuija; Koivula, Teija; Jestoi, M.; Rizzo, A.; Paavanen-Huhtala, S.; Yli-Mattila, T.; Haikara, Auli.

2004. FOSARE Seminar Series 5 Contaminants and Influence of Agricultural Practices, Brussels, Belgium.

Research output: Contribution to conferenceOther conference contributionScientific

TY - CONF

T1 - Real-time PCR for quantification of toxigenic Fusarium species in cereals

AU - Sarlin, Tuija

AU - Koivula, Teija

AU - Jestoi, M.

AU - Rizzo, A.

AU - Paavanen-Huhtala, S.

AU - Yli-Mattila, T.

AU - Haikara, Auli

PY - 2004

Y1 - 2004

N2 - Fusarium species are potential mycotoxin producers in cereals. In near future EU is going to set the maximum limit values for Fusarium toxins in raw cereals and cereal products. Mycotoxin analyses are expensive and time-consuming hence a rapid and reliable quantification method for toxigenic Fusarium species is needed in order to be able to evaluate the mycotoxin risk in cereal-based industry. We have applied real-time PCR technique for the quantification of trichothecene-producing Fusarium species present in barley and malt samples. PCR results were compared to the amount of trichothecenes determined in the samples. Furthermore, highly toxigenic Fusarium graminearum was quantified in cereals by real-time PCR. DNA was extracted from ground kernels using FastDNA Spin Kit for Soil and analysed in a LightCycler„¥ system using the double-stranded DNA ¡Vbinding fluorescent dye SYBR Green I or the fluorigenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. PCR results correlated with the trichothecene content of the grain samples. Non-specific products such as primer dimers were amplified in the SYBR Green I assay, which interfered the quantification. Non-specific amplicons were not detected with the TaqMan probes. In addition, this assay was regarded as sensitive and reproducible.

AB - Fusarium species are potential mycotoxin producers in cereals. In near future EU is going to set the maximum limit values for Fusarium toxins in raw cereals and cereal products. Mycotoxin analyses are expensive and time-consuming hence a rapid and reliable quantification method for toxigenic Fusarium species is needed in order to be able to evaluate the mycotoxin risk in cereal-based industry. We have applied real-time PCR technique for the quantification of trichothecene-producing Fusarium species present in barley and malt samples. PCR results were compared to the amount of trichothecenes determined in the samples. Furthermore, highly toxigenic Fusarium graminearum was quantified in cereals by real-time PCR. DNA was extracted from ground kernels using FastDNA Spin Kit for Soil and analysed in a LightCycler„¥ system using the double-stranded DNA ¡Vbinding fluorescent dye SYBR Green I or the fluorigenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. PCR results correlated with the trichothecene content of the grain samples. Non-specific products such as primer dimers were amplified in the SYBR Green I assay, which interfered the quantification. Non-specific amplicons were not detected with the TaqMan probes. In addition, this assay was regarded as sensitive and reproducible.

M3 - Other conference contribution

ER -

Sarlin T, Koivula T, Jestoi M, Rizzo A, Paavanen-Huhtala S, Yli-Mattila T et al. Real-time PCR for quantification of toxigenic Fusarium species in cereals. 2004. FOSARE Seminar Series 5 Contaminants and Influence of Agricultural Practices, Brussels, Belgium.