DNA diagnostics based on mass spectrometry

{
  "publication_number": "US6221605B1",
  "country": "US",
  "kind": "B1",
  "title": "DNA diagnostics based on mass spectrometry",
  "abstract": "Fast and highly accurate mass spectrometry-based processes for detecting particular nucleic acid molecules and sequences in the molecules are provided. Depending upon the sequence to be detected, the processes, for example, can be used to diagnose a genetic disease or a chromosomal abnormality, a predisposition to a disease or condition, or infection by a pathogen, or for determining identity or heredity.",
  "claims": [
    "1. A process for detecting a target nucleic acid sequence present in a biological sample, comprising the steps of: a) obtaining a target nucleic acid sequence from a biological sample; b) replicating the target nucleic acid sequence, thereby producing a replicated molecule; c) specifically digesting the replicated nucleic acid molecule using at least one appropriate nuclease, thereby producing digested fragments; d) immobilizing the digested fragments onto a solid support containing complementary capture nucleic acid sequences to produce immobilized fragments; and e) analyzing the immobilized fragments by mass spectrometry, wherein the determination of the molecular weight of the immobilized fragments provide information on the target nucleic acid sequence.",
    "2. A process for detecting one or more target nucleic acids in a biological sample, comprising: a) digesting one or more nucleic acids using at least one nuclease, thereby producing digested fragments; b) analyzing the digested fragments by mass spectrometry, whereby detection of the target nucleic acid by mass spectrometry indicates the presence of the target nucleic acid sequence in the biological sample.",
    "3. The process claim 2, further comprising the step of amplifying one or more target nucleic acids.",
    "4. The method of claim 2, wherein the digested fragments are conditioned prior to mass spectrometric analysis.",
    "5. The method of claim 2, wherein the nuclease is a restriction endonuclease.",
    "6. The process of claim 2, wherein identification of a target nucleic acid in the sample provides a genetic diagnosis, detects chromosomal aneuploidy, detects a genetic predisposition to a disease or condition, or detects or identifies infection by a pathogen.",
    "7. The process of claim 2, wherein a plurality of nucleic acids from the sample are immobilized on a solid support prior to mass spectrometric analysis.",
    "8. The method of claim 7, wherein the nucleic acids are arranged in an array and each spot on the array is subjected to mass spectrometric analysis.",
    "9. The process of claim 7, wherein immobilization is effected by a bond cleavable by a pyrophosphatase.",
    "10. The process of claim 2, wherein a nucleic acid comprising the target sequence has been contacted with an alkylating agent prior to mass spectrometric analysis.",
    "11. The process of claim 2, wherein a nucleic acid comprising the target sequence includes one or more of the following: nucleotides that reduce sensitivity for depurination, RNA building blocks, phosphorothioate groups, nucleic acid mimetics and protein nucleic acid (PNA).",
    "12. The process of claim 11, wherein the target nucleic acid includes an alkylated phosphorothioate group.",
    "13. A process of claim 1, wherein the solid support is selected from the group consisting of beads, flat surfaces, pins, combs and wafers."
  ],
  "description_excerpt": "The genetic information of all living organisms (e.g. animals, plants and microorganisms) is encoded in deoxyribonucleic acid (DNA). In humans, the complete genome is comprised of about 100,000 genes located on 24 chromosomes (The Human Genome, T. Strachan, BIOS Scientific Publishers, 1992). Each gene codes for a specific protein which after its expression via transcription and translation, fulfills a specific biochemical function within a living cell. Changes in a DNA sequence are known as mutations and can result in proteins with altered or in some cases even lost biochemical activities; this in turn can cause genetic disease. Mutations include nucleotide deletions, insertions or alterations (i.e. point mutations). Point mutations can be either “missense”, resulting in a change in the amino acid sequence of a protein or “nonsense” coding for a stop codon and thereby leading to a truncated protein.\n\nMore than 3000 genetic diseases are currently known (Human Genome Mutations, D. N. Cooper and M. Krawczak, BIOS Publishers, 1993), including hemophilias, thalassemias, Duchenne Muscular Dystrophy (DMD), Huntington's Disease (HD), Alzheimer's Disease and Cystic Fibrosis (CF). In addition to mutated genes, which result in genetic disease, certain birth defects are the result of chromosomal abnormalities such as Trisomy 21 (Down's Syndrome), Trisomy 13 (Patau Syndrome), Trisomy 18 (Edward's Syndrome), Monosomy X (Turner's Syndrome) and other sex chromosome aneuploidies such as Klinefetter's Syndrome (XXY).",
  "cpc": [
    "C12Q 1/6872",
    "C12Q 1/6816",
    "C12Q 1/6827",
    "C12Q 1/6837",
    "C12Q 1/6858",
    "C12Q 1/686",
    "C12Q 1/6862",
    "C12Q 1/6883",
    "C12Q 1/706",
    "G01N 35/1067",
    "H01J 49/00",
    "Y10T 436/143333",
    "Y10T 436/24"
  ],
  "ipc": [
    "C12N 15/09",
    "C12Q 1/68",
    "C12Q 1/70",
    "G01N 27/62",
    "G01N 33/50"
  ],
  "assignees": [
    "Sequenom Inc"
  ],
  "inventors": [
    "Hubert Köster"
  ],
  "filing_date": "2000-02-15",
  "publication_date": "2001-04-24",
  "grant_date": "2001-04-24",
  "priority_date": "1995-03-17",
  "application_number": "US-50424500-A",
  "family_id": "23606950",
  "cited_by_count": 210,
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