Method for detecting nucleic acid sequences using...

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2223823

The present invention provides a method for quantitatively detecting the amount of a target nucleic acid sequence which may be present in a test sample. A test sample which may contain a target nucleic acid sequence comprising target sequences X and Y is contacted with two primer sets, the first set being specific for target X and the second set being specific for target Y. The test sample also is contacted at the same time with an internal standard sequence IS, which is substantially derived from a combination of the first and second target sequences, and its corresponding oligonucleotide primers. Haptens are associated with the oligonucleotide primer sets in such a way that amplified target sequence products X and Y are detected by capture on a solid phase to which anti-hapten capture reagents are attached. A signal ratio of (X + Y)/S is determined to quantitate the amount of the target nucleic acid sequence contained in the sample.

L'invention concerne une méthode pour la détection quantitative de la quantité d'une séquence d'acide nucléique cible pouvant être présente dans un échantillon test. Un échantillon test pouvant contenir une séquence d'acide nucléique cible comprenant des séquences cibles X et Y est mis en contact avec deux ensembles amorces, le premier ensemble étant spécifique à la cible X et le second ensemble étant spécifique à la cible Y. L'échantillon test est également mis en contact en même temps avec une séquence standard interne IS laquelle est dérivée d'une combinaison des première et seconde séquences cibles, et ses amorces oligonucléotides correspondantes. Des haptènes sont associés aux ensembles amorces oligonucléotidiques de telle manière que les produits des séquences cibles amplifiés X et Y sont détectés par capture sur une phase solide sur laquelle les réactifs de capture d'anti-haptènes sont fixés.Un rapport de signal (X + Y)/S est déterminé pour quantifier la séquence d'acide nucléique cible contenue dans l'échantillon.

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