Method for the simultaneous detection of multiple nucleic...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2692633

The invention is in the technical field of detecting nucleic acid sequences in a sample, such as the detection of pathogenic organisms in clinical samples. More specifically, the invention relates to the field of detecting an infection caused by a pathogenic organism such as a virus or a bacterium in a clinical specimen by means of amplifying and detecting specific nucleic acid sequences from said pathogenic organism. It provides a multiplex assay with the possibility to determine about 30 different target nucleic acid sequences in a single one-tube assay combined with real- time probe detection. The method employs multiplex ligation dependent Probe Amplification (MLPA) employing labelled primers in combination with labelled, probes and detection of the fluorescence.

La présente invention concerne la détection de séquences d'acides nucléiques dans un échantillon, et notamment la détection d'organismes pathogènes dans des échantillons cliniques. L'invention concerne plus particulièrement la détection d'une infection causée par un organisme pathogène tel qu'un virus ou une bactérie dans un spécimen clinique au moyen de l'amplification et de la détection de séquences d'acides nucléiques spécifiques dudit organisme pathogène. L'invention concerne un test multiplex avec la possibilité de déterminer environ 30 séquences d'acides nucléiques cibles différentes dans un test mono-tube unique combiné avec une détection par sonde en temps réel. Le procédé utilise l'amplification multiplex de sonde nucléique dépendant des ligatures (MLPA) utilisant des amorces marquées en association avec des sondes marquées et la détection de la fluorescence.

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