Colorimetric genetic test for clinically significant TNF polymorphism and methods of use thereof
OwnerYi-Wei Tanginventor
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Abstract
The present invention discloses a method and diagnostic kit used to identify polymorphisms. The method makes use of a minor groove binder and shorter hybridization probes so that polymorphisms, including polymorphisms of only a single nucleotide, may be detected. The present invention additionally discloses methods using hybridization conditions which increase the sensitivity of each individual nucleotide within the region being assayed. The invention additionally includes a diagnostic kit which supplies the necessary materials as well as instructions so that the method disclosed herein may be practiced in a time efficient manner.
Claims30
A method of identifying a nucleotide variation in sequence in a nucleic acid, comprising:
- providing a biological sample having a nucleic acid;
- contacting the nucleic acid with a primer under conditions for nucleic acid amplification, wherein the primer amplifies the nucleic acid in order to produce an amplification product, wherein the nucleic acid is amplified by polymerase chain reaction;
- contacting a first aliquot of the amplification product with a first solution having a first biotin-labeled probe, the first biotin-labeled probe having a minor groove binder so that an amplification product-probe complex is formed;
- and
- detecting the amplification product-probe complex.
A method of identifying a genetic predisposition in a mammal, comprising:
- providing a biological sample having genomic DNA from the mammal, wherein the genomic DNA is tumor necrosis factor alpha;
- amplifying the genomic DNA in order to produce an amplification product;
- providing a hybridization solution so that a probe hybridizes to the amplification product;
- hybridizing the probe to the amplification product so that an amplification product-probe complex is formed;
- immobilizing the probe;
- and
- determining the presence of the amplification product-probe complex, wherein the presence of the amplification product-probe complex identifies a single nucleotide polymorphism in the tumor necrosis factor alpha genomic DNA.
A diagnostic kit for detecting single nucleotide polymorphisms in mammals, comprising:
- a forward polymerase chain reaction primer;
- a reverse polymerase chain reaction primer;
- a first probe having a first minor groove binding component;
- a second probe having a second minor groove binding component;
- and
- a set of instructions for use.
Dependencies are inferred from the claim text.