Apparatus and method for detecting genetic mutations and single nucleotide polymorphisms
A method and means of identifying nucleic acid oligomers is disclosed. A sample is split into parts and the parts are flowed through chromatography columns containing nucleic acid oligomer probes bound to a binding medium. Analyte oligomers are transiently hybridized to complementary probe oligomers during chromatography. Detection and analysis of oligomer peaks is used to identify the oligomers contained in the sample.
1. A method for identifying nucleic acid mutations comprising;
obtaining a sample of target nucleic acid oligomers comprising at least one target sequence;
where the target sequence is also known to exist in mutated form;
dividing the sample into at least two parts;
loading the parts onto at least two portions of binding medium;
where each part is loaded onto a separate portion of binding medium;
where the binding medium comprises probe nucleic acid oligomers bound to a solid support;
where a first portion of binding medium comprises probe oligomers that are complementary to the at least one target sequence;
where a second portion of binding medium comprises probe oligomers that are complementary to a mutated form of the at least one target sequence;
where the rates of hybridization and dissociation of the target oligomers and the probe oligomers is about the same;
flowing the oligomer-containing sample parts through the binding medium portions;
detecting oligomer peaks in the fluid exiting from each of the at least two portions of binding medium;
analyzing the oligomer peak data from the at least two portions of binding medium to identify the target nucleic acid oligomer mutations.
2. The method of claim 1 further comprising;
dividing the sample into at least a third part;
loading the at least a third part onto at least a third portion of binding medium;
where the at least a third portion of binding medium comprises probe oligomers that are complementary to a sequence different from the at least one target sequence and differing from the mutated form of the at least one target sequence.
3. The method of claim 1 wherein the ratio of the rate of hybridization to the rate of dissociation is between about 35:65 and about 65:35.
4. The method of claim 1 further comprising the use of an internal standard.
5. The method of claim 1 where probe oligomers are covalently attached to the binding medium.
6. The method of claim 1 where probe oligomers are non-covalently attached to the binding medium.
7. The method of claim 1 where target oligomers comprise DNA.
8. The method of claim 1 where target oligomers further comprise a label.
9. The method of claim 1 where the at least one target sequence is associated with a human genetic disease.
10. The method of claim 9 where the disease is glucose-6-phosphate dehydrogenase deficiency.
11. The method of claim 9 where the disease is cystic fibrosis.
12. The method of claim 9 where the disease is muscular dystrophy.
13. The method of claim 9 where the disease is breast cancer.
14. The method of claim 9 where the disease is familial hypercholesterolemia.
15. The method of claim 9 where the disease is sickle cell anemia.