EFS12- Book of abstracts - Contact
EFS12- Book of abstracts - Contact
EFS12- Book of abstracts - Contact
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SESSION 3: PATHOGENESIS – EPIDEMIOLOGY AND POPULATION<br />
GENETICS<br />
P71 - Real time PCR for FHB quantification: bias<br />
analysis?<br />
S. Elbelt 1 , S. Fromentin 1 , V. Laval 1 , C. Barreau 2 , F. Carpentier 1 , C. Ducos 2 , C.<br />
Lannou 1 , F. Techini 1 , J. Toussaint 3 , C. Vitry 3 , L. Pinson-Gadais 2<br />
1 INRA, UR1290 Bioger_CPP, bat 13, avenue L. Bretignière, F-78850 Thiverval Grignon, France;<br />
2 INRA, UR1264 MycSA, 71 Avenue Edouard Bourlaux, CS 20032, F-33882 Villenave d’Ornon,<br />
France; 3ARVALIS Institut du Végétal, bat 13, avenue L. Bretignière, F-78850 Thiverval Grignon,<br />
France<br />
E-mail: vlaval@versailles.inra.fr<br />
A better understanding <strong>of</strong> the FHB species complex and particularly <strong>of</strong> the way<br />
species interactions influence epidemic development and toxin production<br />
requires accurate, sensitive and specific molecular diagnostic tools, such as realtime<br />
PCR. Nine qPCR tests specifically designed for the most represented<br />
species <strong>of</strong> the FHB complex on wheat in Europe: F. graminearum, F. culmorum,<br />
F. poae, F. avenaceum, F. langsethiae, F. tricinctum, F. sporotrichioides, M.<br />
majus and M. nivale were assessed for their potential for diagnosis. We present<br />
here the results <strong>of</strong> quantification analysis performed in three laboratories which<br />
use different mix and real time PCR apparatus. The variation in quantification will<br />
be discussed. In one laboratory the limits <strong>of</strong> the diagnostic was further studied by<br />
analysing DNA extraction and real time PCR repeatability. Statistical analysis was<br />
performed on results obtained from two extractions <strong>of</strong> 222 samples and three real<br />
time PCR analysis performed per extraction. First results show the importance to<br />
carefully test real time PCR parameters when developing real time PCR tools for<br />
diagnosis or epidemiological studies.<br />
Keywords: Molecular diagnostic tools, real-time PCR, FHB, inter-laboratory<br />
validation tests<br />
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