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Nucleic acid assays

  • US 7,262,008 B2
  • Filed: 08/08/2002
  • Issued: 08/28/2007
  • Est. Priority Date: 05/02/1997
  • Status: Expired due to Term
First Claim
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1. A method for the automated detection of the presence or absence of a first target nucleic acid and a second target nucleic acid in a sample, wherein the first target nucleic acid is different from the second target nucleic acid, said method comprising:

  • a) contacting said sample with a reaction vessel, wherein said reaction vessel is coated with a first capture nucleic acid which can form a specific hybridization complex with said first target nucleic acid, and said reaction vessel is coated with a second capture nucleic acid which can form a specific hybridization complex with said second target nucleic acid, wherein the first capture nucleic acid is different from the second capture nucleic acid;

    b) allowing specific hybridization complexes to form if said first target nucleic acid or said second target nucleic acid or both is in said sample;

    c) contacting said specific hybridization complexes with a first detection nucleic acid, wherein said first detection nucleic acid can form a specific hybridization detection complex with said first target nucleic acid, and wherein said first detection nucleic acid is conjugated to one or more compounds selected from the group consisting of enzyme, chromophore, chemiluminescent compound, radioisotope, and fluorophore;

    d) allowing first specific detection complexes to form if said first target nucleic acid is in said sample;

    e) detecting one or more detectable signals generated from the one or more compounds of step c) if said first target nucleic acid is in said sample;

    f) contacting said specific hybridization complexes with a second detection nucleic acid, wherein the second detection nucleic acid is different from the first detection nucleic acid, and wherein said second detection nucleic acid can form a specific hybridization detection complex with said second target nucleic acid, and wherein said second detection nucleic acid is conjugated to one or more compounds selected from the group consisting of enzyme, chromophore, chemiluminescent compound, radioisotope, and fluorophore;

    g) allowing second specific detection complexes to form if said second target nucleic acid is in said sample; and

    h) detecting one or more detectable signals generated from the compound of step f) if said second target nucleic acid is in said sample;

    wherein the absence of a detectable signal correlates with the absence of said first target nucleic acid and said second target nucleic acid in said sample and the presence of said one or more detectable signals in step e) or step h) indicates the presence of said first target nucleic acid or the presence of said second target nucleic acids, respectively in said sample and wherein optionaly, between step b) and c) or d) and e) or g) and h), said specific hybridization complex are washed.

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