EFS12- Book of abstracts - Contact

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