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Detection of microbial nucleic acids by a one-step sandwich hybridization test

  • US 4,486,539 A
  • Filed: 10/14/1982
  • Issued: 12/04/1984
  • Est. Priority Date: 10/16/1981
  • Status: Expired due to Term
First Claim
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1. A method for identifying nucleic acids by a one-step sandwich hybridization test, said method comprising the steps of:

  • (a) rendering the nucleic acids in the sample to be identified single-stranded;

    (b) allowing said single-stranded nucleic acids of the sample to hybridize simultaneously in a single step with a combination of at least one pair of nucleic acid reagents, said reagents having been purified, the first nucleic acid reagent of said pair comprising a single-stranded fragment of nucleic acid, having a nucleotide sequence of at least 10 bases, and being affixed to a solid carrier, the second nucleic acid reagent of said pair comprising a single-stranded fragment of nucleic acid, having a nucleotide sequence of at least 10 bases, and being labeled with a radioisotope, said first and second nucleic acid reagents being capable of forming hybrid molecules by complementary base pairing with given sequences of the sample nucleic acid to be identified, provided that the second nucleic acid reagent cannot hybridize with the first nucleic acid reagent;

    (c) washing said solid carrier to substantially remove said label which is not incorporated in said hybrid molecule; and

    (d) measuring said label on the washed solid carrier, whereby determining whether the sample contains the nucleic acid to be identified.

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