Rapid genotyping analysis and the device thereof
AssigneeDiagCor Bioscience Incorporation Ltd.
InventorJoseph Wing On Tam
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Abstract
The present invention discloses the use of Allele-Specific-Oligonucleotide (ASO) as a detection assay for human HLA classification. Using Reversed-Dot-Blotting format and flow through hybridization process, more efficient, faster and less expensive HLA classification can be achieved. A simplified procedure for HLA genotyping is also described. This invention further provides a Single Nucleotide Polymorphism (SNP)-based DNA fingerprining method for rapid and accurate genotyping, identification as well as DNA analyses of genetic data from human beings and different organisms. In addition this invention also discloses a new device for rapid and sensitive analyses of nucleic acids, proteins and other analysts for diagnosis.
Claims16
A method for rapid identification of HLA Class II genotype of a subject, comprising the steps of:
- (a) obtaining a sample comprising a template nucleic acid from the subject;
- (b) amplifying the template nucleic acid with primers having the sequences of SEQ ID NOs 97-106, thereby generating HLA amplicons;
- and
- (c) hybridizing the HLA amplicons with immobilized oligonucleotide probes having the sequences of SEQ ID NOs 1, 2, 4, 5, 7, 8, 10, 12-15, 17, 19-21, 24, 26, 29-55, 57-64, 66, 67, 69-88, and 90-92 wherein the resulting hybridization profile would indicate the HLA Class II genotype of the subject.
A method of rapid DNA fingerprinting of a subject, comprising the steps of:
- a) obtaining a sample comprising template nucleic acid molecules from the subject;
- b) amplifying the template nucleic acid molecules with primers having the sequences of SEQ ID NOs 97-106, wherein the primers flank one or more loci of interest and comprise signal generating labels;
- c) immobilizing oligonucleotide capture probes on a membrane, wherein the probes have the sequences of SEQ ID NOs 1, 2, 4, 5, 7, 8, 10, 12-15, 17, 19-21, 24, 26, 29-55, 57-64, 66, 67, 69-88, and 90-92;
- d) applying the amplified nucleic acid molecules in a hybridization solution to the membrane without pre-hybridization;
- and
- e) hybridizing the amplified nucleic acid molecules with the oligonucleotide capture probes, wherein the hybridization is carried out in a lateral flow-through process wherein sensitivity of the hybridization depends on the ratio of the area comprising the probes to the cross section of the membrane.
Dependencies are inferred from the claim text.