Method for the sensitive detection of nucleic acids

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 5, 435 912, 435174, 536 231, 536 265, 536 243, 536 2432, 536 2433, C12Q 168, C12Q 170, C12P 1934, C07H 2104

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057534330

ABSTRACT:
Method for the specific detection of nucleic acids in a sample by reaction of the sample with one or several labelled nucleotide triphosphates and one or several enzymes which catalyse the production of a labelled nucleic acid B which contains this nucleotide, non-thermally denaturation, reacting the sample with a nucleic acid probe C which is sufficiently complementary to nucleic acid B and which contains at least one immobilizable group, if desired, contacting the nucleic acid hybrid D formed with a solid phase which recognizes and binds the immobilizable group, removing the liquid from the solid phase and determining the label on the solid phase as a measure for the presence of the nucleic acid.

REFERENCES:
patent: 5200314 (1993-04-01), Urdea et al.
patent: 5232829 (1993-08-01), Longiarn et al.
Proc. Natl. Acad. Sci. USA; 87, pp. 1874-1878 (1990) Isothermal, in-vitro amplification of nucleic acids by a multienzyme reaction modeled after retroviral replication.
Proc. Natl. Acad. Sci. USA; 86, pp. 6230-6234 (1989) Genetic analysis of amplified DNA with immobilized sequence-specific oligonucleotide probes.
Proc. Natl. Acad. Sci. USA 86, pp. 1173-1177 (1989) Transcription-based amplification system and detection of amplified human immunomodeficiency virus type 1 with a bead-based sandwich hybridization format.
Innis and Gelfand, in PCR Protocols, A Guide to Methods and Applications, pp. 3-11, Academic Press, 1990.

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