MASS SPECTROMETRIC METHODS FOR DETECTING MUTATIONS IN A TARGET NUCLEIC ACID
Fast and highly accurate mass spectrometry-based processes for detecting particular nucleic acid molecules and mutations in the molecules are provided.
1 . A process for detecting one or more target nucleotide sequences in a biological sample, comprising:
a) amplifying a target nucleic acid molecule, wherein the amplification product is a natural deoxyribonucleic acid;
b) ionizing and volatilizing the product of step (a); and
c) analyzing the product of step (b) by mass spectrometry, wherein detection of the target nucleotide sequence by mass spectrometry indicates the presence of the target nucleotide sequence in the biological sample.
2 . The process of claim 1 , wherein the target nucleotide sequence is a mutation.
3 . The process of claim 1 , wherein a plurality of nucleic acids are arranged on a solid support in an array and each spot on the array is subject to mass spectrometry.
4 . The process of claim 1 , wherein the mass spectrometry format is matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry.
5 . The process of claim 1 , wherein the mass spectrometry format is electrospray (ES) mass spectrometry.
6 . The process of claim 1 , wherein the target nucleotide sequence is a DNA fingerprint or is implicated in a disease or condition selected from the group consisting of a genetic disease, a chromosomal abnormality, a genetic predisposition, a viral infection, a fungal infection, a bacterial infection and a protest infection.
7 . The process of claim 6 , whereby a genetic disease, chromosomal abnormality, genetic predisposition to a disease or condition or infection by a pathogen can be detected or identity determined.
8 . The process of claim 1 , which further comprises exchanging cations bound to the product of step (a).
9 . The process of claim 1 , wherein the product of step (a) and step (b) are natural oligoribo nucleic acids or natural oligodeoxyribo nucleic acids.
10 . The process of claim 1 , wherein the backbone of the product of step (a) and step (b) is a phosphodiester backbone.
11 . The process of claim 1 , wherein each sugar-sugar linkage in the target nucleic acid comprises a negative charge.
12 . The process of claim 1 , wherein each sugar-sugar linkage in the amplification product comprises a negative charge.
13 . The process of claim 1 , wherein each sugar-sugar linkage in the target nucleic acid comprises a phosphate.
14 . The process of claim 1 , wherein each sugar-sugar linkage in the amplification product comprises a phosphate.
15 . The process of claim 1 , wherein the product of step (a) is contacted with a capture oligonucleotide linked to a solid support.
16 . The process of claim 1 , wherein the target nucleic acid is contacted with a capture oligonucleotide linked to a solid support.