Real-time PCR for quantification of toxigenic Fusarium species in barley and malt

Tuija Sarlin, 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 some Fusarium toxins in unprocessed cereals and cereal products. Mycotoxin analyses are expensive and time-consuming. Hence, a rapid and reliable quantification method for toxigenic Fusarium species is needed for evaluation of 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 (the TMTRI assay, S. Klemsdal unpubl. sequences). PCR results were compared to the amount of trichothecenes in the samples. Furthermore, highly toxigenic Fusarium gram inearum was quantified in cereals by real-time PCR (the TMFg12 assay, T. Yli-Mattila unpubl. sequences). DNA was extracted from ground kernels (0.1 g) using FastDNA Spin Kit for Soil and analysed in a LightCycler® system using fluorigenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium species and F. graminearum present in barley and malt samples, respectively. Both TaqMan assays were regarded as sensitive and reproducible. Linearity of the assays was at least 3-4 log units when determined using pure Fusarium DNA. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay correlated with the DON content in Finnish barley samples. The TMFg12 assay for F. graminearum gave a good estimation about the DON content in North American barley and malt samples. The amounts of DON and F. graminearum in Finnish barley were found to be naturally low.
Original languageEnglish
Publication statusPublished - 2004
MoE publication typeNot Eligible
Event2nd International Symposium on Fusarium Head Blight - Orlando, United States
Duration: 11 Dec 200415 Dec 2004

Conference

Conference2nd International Symposium on Fusarium Head Blight
CountryUnited States
CityOrlando
Period11/12/0415/12/04

Fingerprint

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

Keywords

  • Fusarium
  • scab
  • proteinases
  • barley
  • cereals

Cite this

Sarlin, T., Jestoi, M., Rizzo, A., Paavanen-Huhtala, S., Yli-Mattila, T., & Haikara, A. (2004). Real-time PCR for quantification of toxigenic Fusarium species in barley and malt. 2nd International Symposium on Fusarium Head Blight, Orlando, United States.
Sarlin, Tuija ; Jestoi, M. ; Rizzo, A. ; Paavanen-Huhtala, S. ; Yli-Mattila, T. ; Haikara, Auli. / Real-time PCR for quantification of toxigenic Fusarium species in barley and malt. 2nd International Symposium on Fusarium Head Blight, Orlando, United States.
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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 some Fusarium toxins in unprocessed cereals and cereal products. Mycotoxin analyses are expensive and time-consuming. Hence, a rapid and reliable quantification method for toxigenic Fusarium species is needed for evaluation of 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 (the TMTRI assay, S. Klemsdal unpubl. sequences). PCR results were compared to the amount of trichothecenes in the samples. Furthermore, highly toxigenic Fusarium gram inearum was quantified in cereals by real-time PCR (the TMFg12 assay, T. Yli-Mattila unpubl. sequences). DNA was extracted from ground kernels (0.1 g) using FastDNA Spin Kit for Soil and analysed in a LightCycler{\circledR} system using fluorigenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium species and F. graminearum present in barley and malt samples, respectively. Both TaqMan assays were regarded as sensitive and reproducible. Linearity of the assays was at least 3-4 log units when determined using pure Fusarium DNA. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay correlated with the DON content in Finnish barley samples. The TMFg12 assay for F. graminearum gave a good estimation about the DON content in North American barley and malt samples. The amounts of DON and F. graminearum in Finnish barley were found to be naturally low.",
keywords = "Fusarium, scab, proteinases, barley, cereals",
author = "Tuija Sarlin and M. Jestoi and A. Rizzo and S. Paavanen-Huhtala and T. Yli-Mattila and Auli Haikara",
year = "2004",
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Sarlin, T, Jestoi, M, Rizzo, A, Paavanen-Huhtala, S, Yli-Mattila, T & Haikara, A 2004, 'Real-time PCR for quantification of toxigenic Fusarium species in barley and malt' 2nd International Symposium on Fusarium Head Blight, Orlando, United States, 11/12/04 - 15/12/04, .

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

2004. 2nd International Symposium on Fusarium Head Blight, Orlando, United States.

Research output: Contribution to conferenceOther conference contributionScientific

TY - CONF

T1 - Real-time PCR for quantification of toxigenic Fusarium species in barley and malt

AU - Sarlin, Tuija

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 some Fusarium toxins in unprocessed cereals and cereal products. Mycotoxin analyses are expensive and time-consuming. Hence, a rapid and reliable quantification method for toxigenic Fusarium species is needed for evaluation of 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 (the TMTRI assay, S. Klemsdal unpubl. sequences). PCR results were compared to the amount of trichothecenes in the samples. Furthermore, highly toxigenic Fusarium gram inearum was quantified in cereals by real-time PCR (the TMFg12 assay, T. Yli-Mattila unpubl. sequences). DNA was extracted from ground kernels (0.1 g) using FastDNA Spin Kit for Soil and analysed in a LightCycler® system using fluorigenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium species and F. graminearum present in barley and malt samples, respectively. Both TaqMan assays were regarded as sensitive and reproducible. Linearity of the assays was at least 3-4 log units when determined using pure Fusarium DNA. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay correlated with the DON content in Finnish barley samples. The TMFg12 assay for F. graminearum gave a good estimation about the DON content in North American barley and malt samples. The amounts of DON and F. graminearum in Finnish barley were found to be naturally low.

AB - Fusarium species are potential mycotoxin producers in cereals. In near future EU is going to set the maximum limit values for some Fusarium toxins in unprocessed cereals and cereal products. Mycotoxin analyses are expensive and time-consuming. Hence, a rapid and reliable quantification method for toxigenic Fusarium species is needed for evaluation of 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 (the TMTRI assay, S. Klemsdal unpubl. sequences). PCR results were compared to the amount of trichothecenes in the samples. Furthermore, highly toxigenic Fusarium gram inearum was quantified in cereals by real-time PCR (the TMFg12 assay, T. Yli-Mattila unpubl. sequences). DNA was extracted from ground kernels (0.1 g) using FastDNA Spin Kit for Soil and analysed in a LightCycler® system using fluorigenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium species and F. graminearum present in barley and malt samples, respectively. Both TaqMan assays were regarded as sensitive and reproducible. Linearity of the assays was at least 3-4 log units when determined using pure Fusarium DNA. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay correlated with the DON content in Finnish barley samples. The TMFg12 assay for F. graminearum gave a good estimation about the DON content in North American barley and malt samples. The amounts of DON and F. graminearum in Finnish barley were found to be naturally low.

KW - Fusarium

KW - scab

KW - proteinases

KW - barley

KW - cereals

M3 - Other conference contribution

ER -

Sarlin T, Jestoi M, Rizzo A, Paavanen-Huhtala S, Yli-Mattila T, Haikara A. Real-time PCR for quantification of toxigenic Fusarium species in barley and malt. 2004. 2nd International Symposium on Fusarium Head Blight, Orlando, United States.