Method for detecting charged oligonucleotides in biological...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C12N 15/10 (2006.01)

Patent

CA 2176341

Disclosed is a method for detecting and quantitating oligonucleotides with charged internucleotide linkages in biological fluids. In this method,'a biological fluid sample is contacted with an anion exchange resin at from 40 °C to 65 °C for a time sufficient to enable oligonucleotides in the sample to adsorb to the resin. The adsorbed oligonucleotides are then desorbed with a buffer having a salt concentration of about 1 M to 2.5 M and a pH in the range of about 6.5 to 7.5, the desorption being performed at about 40 - 65 °C. The oligonucleotides so released are then detected and quantitated.

On décrit un procédé qui permet de détecter et quantifier des oligonucléotides, à liaisons inter-nucléotidiques chargées, dans des fluides biologiques. Il consiste à mettre un échantillon d'un fluide biologique en contact avec une résine d'échange anionique, entre 40 et 65 ~C, pendant une durée suffisante pour que des oligonucléotides présents dans cet échantillon s'adsorbent dans cette résine. Les oligonucléotides adsorbés sont alors désorbés avec un tampon à concentration en sels allant de 1 à 2,5 M et à pH variant entre 6,5 et 7,5 environ, cette désorption intervenant entre 40 et 65 ~C environ. Les oligonucléotides ainsi libérés font alors l'objet d'une détection et d'une quantification.

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