Method of in-vitro detection and quantification of hiv dna...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01)

Patent

CA 2573591

Process for the detection and quantification in-vitro of HIV DNA by quantitative PCR. The invention consists of developing pairs of oligonucleotides capable of hybridizing with fragments of the gag gene sequence present in the genome of the HIV virus. These oligonucleotides permit amplification of the viral DNA by quantitative PCR. Even in the case of samples with low viral load, the invention permits detection of the HIV DNA virus by juxtaposition either of a conventional PCR with a quantitative PCR, or with a double nested quantitative PCR, using different pairs of primers in each amplification. The invention, compared with the known methods, allows determining qualitatively and quantitatively, in-vitro, the presence of HIV DNA in samples, in a rapid, reproducible manner and with high sensitivity.

La présente invention concerne un procédé de détection et numération in-vitro de l'ADN du VIH par PCR (amplification par polymérase) quantitative. L'invention consiste à développer des paires d'oligonucléotides capables de s'hybrider avec des fragments de la séquence du gènes "gag" présents dans le génome du VIH. Ces oligonucléotides permettent l'amplification de l'ADN viral par PCR quantitative. Même dans le cas d'échantillons à faible charge virale, l'invention permet la détection du virus à ADN de VIH par juxtaposition soit d'une PCR conventionnelle avec une PCR quantitative, soit avec une PCR quantitative imbriquée double, au moyen de paires différentes d'amorces dans chaque amplification. Par rapport aux procédés connus, l'invention permet une détermination qualitative et quantitative in-vitro de la présence d'ADN du VIH d'une façon rapide et reproductible, avec un niveau élevé de sensibilité.

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