Sample preparation for nucleic acid based diagnostic tests

C - Chemistry – Metallurgy – 12 – Q

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Details

C12Q 1/68 (2006.01) C12N 15/09 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 2236613

The present invention provides a method for increasing the sensitivity of a diagnostic test for the detection of a nucleic acid of an infectious agent in pooled samples of liquid biological material, characterized in that the nucleic acid is selectively precipitated by adding a precipitating agent, whereby the preferred precipitating agent is polyethylene glycol in a molecular weight range of 1'000 to 10'000 at a final concentration of 1 to 5 % (w/v). The precipitation efficiency may be increased by the addition of mono- or divalent salts. Preferred examples of such salts are NaCl or KCl, preferably these salts are added so that the final salt concentration is 200 to 450 mM, most preferably 270 to 325 mM salt. The invention also relates also to the use of the said method in a diagnostic test as well as to kits which are useful for practicing the said method.

Cette invention fournit une méthode pour rehausser la sensibilité d'un test diagnostique permettant de détecter l'acide nucléique d'un agent infectieux dans des échantillons mis en pool de matériel biologique liquide, caractérisée par le fait que l'acide nucléique est précipité de façon sélective par l'ajout d'un agent de précipitation, où l'agent de précipitation privilégié est du polyéthylèneglycol dont le poids moléculaire va de 1000 à 10 000 à une concentration finale de 1 à 5 % (p/v). On peut accroître l'efficacité de la précipitation en ajoutant des sels monovalents ou bivalents. Des exemples privilégiés de ces sels sont NaCl et KCl, ces sels étant de préférence ajoutés de manière à ce que la concentration finale en sel soit de 200 à 450 mM, et préférablement entre 270 et 325 mM. L'invention porte également sur l'utilisation de ladite méthode dans un test diagnostique ainsi que dans des trousses utiles pour l'application de ladite méthode.

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