Method for the detection of fusarium graminearum

Details Method for the detection of fusarium graminearum Method for the detection of fusarium graminearum

C - Chemistry, Metallurgy

12

Q

C12Q 1/68 (2006.01)

Patent

CA 2569190

A method for the detection of Fusarium graminearum (Gibberella zeae) comprising the steps: providing a sample containing a nucleic acid, contacting said sample with at least one forward primer and at least one reverse primer, wherein the at least one reverse primer hybridises within the ~-tubulin nucleic acid sequence of Fusarium graminearum (Gibberella zeae) and comprises the nucleic acid sequence 5'-R1TTTTCGTX1GX2AGT-3' (SEQ ID NO 1), wherein R1 comprises at least one nucleic acid residue of the ~-tubulin nucleic acid sequence located upstream of the hybridisation site of the nucleic acid sequence SEQ ID NO 1 and subsequent to the nucleic acid sequence SEQ ID NO 1, X1 is guanine, adenine or inosine, X2 is cytosine, adenine or inosine, and wherein the at least one forward primer hybridises upstream of the hybridisation site of a nucleic acid sequence complementary to the at least one reverse primer, subjecting the sample contacted with the at least one forward primer and the at least one reverse primer to a nucleic acid amplification technique, and optionally determing the presence of Fusarium graminearum (Gibberella zeae) in a sample by detecting a nucleic acid amplification product.

L'invention porte sur un procédé de détection du Fusarium graminearum (Gibberella zeae) comportant les étapes suivantes: (i) obtenir un échantillon contenant un acide nucléique; (ii) mettre en contact ledit échantillon avec au moins une amorce sens et une amorce antisens, étant donné que: (a) l'amorce antisens, qui s'hybride dans la séquence d'acides nucléiques de la ß-tubuline de la Fusarium graminearum (Gibberella zeae) comporte la séquence d'acides nucléiques 5'-R1TTTTCGTX1GX2AGT-3' (SEQ ID NO 1), dans laquelle: R1 comporte au moins un résidu d'acide nucléique de la séquence d'acides nucléiques de la ß-tubuline située en amont du site d'hybridation de la séquence d'acides nucléiques SEQ ID NO 1; la séquence suivant la séquence d'acides nucléiques SEQ ID NO 1 est guanine, adénine ou inosine; et X2 est cytosine, adénine ou inosine, et que (b) l'amorce sens, s'hybride en amont du site d'hybridation de la séquence d'acides nucléiques complémentaire de l'amorce antisens; (iii) soumettre ledit échantillon ainsi mis en contact à une technique d'amplification d'acides nucléiques; (iv) et facultativement déterminer la présence de Fusarium graminearum (Gibberella zeae) dans l'échantillon en y détectant un produit d'amplification d'acide nucléique.

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