Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1991-06-06
1994-04-26
Wax, Robert A.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 231, 536 235, 536 241, 536 242, 536 232, 935 8, 935 17, 935 78, C12Q 168, C07H 2104, C12N 1511
Patent
active
053066162
DESCRIPTION:
DESCRIPTION
DETAILED DESCRIPTION OF THE INVENTION
It will be readily apparent to one skilled in the art that various substitutions and modifications may be made to the invention disclosed herein without departing from the scope and the spirit of the invention.
As used herein the term "differentially labeled" indicates that each extension product can be distinguished from all others because it has a different label attached and/or is of a different size and/or binds a specifically labeled oligonucleotide. One skilled in the art would recognize a variety of labels are available. For example these can include radioisotopes, fluorescers, chemluminescers, stains, enzymes and antibodies. Various factors affect the choice of the label. These include the effect of the label on the rate of hybridization and binding of the primer to the DNA, the sensitivity of the label, the ease of making the label, primer, probe or extension of products, the ability to automate the available instrumentation, convenience and the like. For example, differential radioisotope labeling could include .sup.32 P, .sup.3 H and .sup.14 C; differential fluorescers labeling could include fluorescein-5-isothiocyanatetetramethylrhodamine-5-(and -6) isothiocyanate, Texas Red and NBD aminohexanoic acid or a mixture of different labels such as radioisotopes, fluorescers and chemluminescers.
Each specific sample to be tested herein for its duplication in the DNA sequence is derived from genomic DNA. The source of the genomic DNA to be tested can be any medical sample. Some examples of medical samples include blood, semen, vaginal swabs, tissue, mouth wash sample, hair and mixture of body fluids.
As used herein the term "polymerase chain reaction" or "PCR" refers to the PCR procedure described in the patents to Mullis, et al., U.S. Pat. Nos. 4,683,195 and 4,683,202. The procedure basically involves: (1) treating extracted DNA to form single-stranded complementary stands; (2) adding a pair of oligonucleotide primers, wherein one primer of the pair is substantially complementary to part of the sequence in the sense strand and the other primer of each pair substantially complementary to a different part of the same sequence in the complementary antisense strand; (3) annealing the paired primers to the complementary sequence; (4) simultaneously extending the annealed primers from the 3, terminus of each primer to synthesize an extension product complementary to the strands annealed to each primer wherein said extension products after separation from their complement serve as templates for the synthesis of an extension product for the other primer of each pair; (5) separating said extension products from said templates to produce single-stranded molecules; and (6) amplifying said single-stranded molecules by repeating at least once said annealing, extending and separating steps.
As used herein "fragment thereof" refers to a partial sequence from the probe or cosmid which still identifies the polymorphism within the CMT locus.
One embodiment of the present invention is a method of detecting CMT disease comprising the step of measuring the presence or absence of a DNA duplication at a gene locus associated with CMT disease.
In a preferred embodiment of the present invention the duplication is measured by the steps of extracting DNA from a sample to be tested, amplifying the extracted DNA, identifying the presence or absence of a DNA duplication in the amplified extension products. The presence of a DNA duplication indicates that the sample came from an individual with CMT disease.
In the preferred embodiment the amplification is by the polymerase chain reaction and the primers are those identified as Sequence ID No. 2 and Sequence ID No. 3.
The CMT disease usually detected by identification of a duplication is that classified as type A. The duplication is identified as three alleles or two copies of one of two allels of a polymorphic (GT).sub.n dinucleotide repeat sequence where n is the number of repeats. At the CMT locus, n is usually 12 to 33. For RM11-GT n is usua
REFERENCES:
Weber, et al. 1990, Nucleic Acids Research 18:4638.
Bedford, M. T., and van Helden, P. D.; A Method to Analyze Allele-Specific Methylation; BioTechniques 9:744-748 (1990).
Chance, P. F., et al; Genetic Linkage and Heterogeneity in Type I Charcot-Marie-Tooth Disease (Hereditary Motor and Sensory Neuropathy Type I); Am. J. Hum. Genet., 47:915-925 (1990).
Devlin, R. H., et al; Partial Gene Duplication Involving Exon-Alu Interchange Results in Lipoprotein Lipase Deficiency; Am. J. Hum. Genet., 46:112-119 (1990).
Fain, P. R., et al; Genetic Analysis of NF1: Identification of Close Flanking Markers on Chromosome 17; Genomics 1:340-345 (1987).
Franco, B., et al; An Mspl RFLPs at the D17S258 locus; Nucleic Acids Research, 18:7196 (1990).
Herrmann, Bernhard G., et al; A Large Inverted Duplication Allows Homologous Recombination between Chromosomes Heterozygous for the Proximal t Complex Inversion; Cell 48, 813-825 (1987).
Kornreich, Ruth, et al; .alpha.-Galactosidase A Gene Rearrangements Causing Fabry Disease; The Journal of Biological Chemistry, 265:9319-9326 (1990).
Lawrence, Jeanne Bentley, et al; Interphase and Metaphase Resolution of Different Distances Within the Human Dystrophin Gene. Science, 249:928-932 (1990).
Litt, Michael; A Hypervariable Microsatellite Revealed by in Vitro Amplification of a Dinucleotide Repeat within the Cardiac Muscle Actin Gene; Am. J. Hum. Genet. 44:397-401 (1989).
Lupski, J. R., et al; Charcot-Marie-Tooth Polyneuropathy Syndrome: Clinical, Electrophysiologic, and Genetic Aspects; In Current Neurology S. Appel, ed. (Chicago, Mosby-Yearbook), pp. 1-25 (1991).
Magenis, R. Ellen, et al; De Novo Partial Duplication of 17p [dup(17)(p12-p11.2)]: Clinical Report; American Journal of Medical Genetics 24:415-420 (1986).
McAlpine, P. J., et al; Localization of a Locus for Charcot-Marie-Tooth Neuropathy Type la (CMT1A) to Chromosome 17; Genomics 7, 408-415 (1990).
Middleton-Price, H. R., et al; Linkage of Hereditary Motor and Sensory Neuropathy Type I to the Pericentromeric Region of Chromosome 17; Am. J. Hum. Genet. 46:92-94, 1990.
Nakamura, Y., et al; A Mapped Set of DNA Markers for Human Chromosome 17; Genomics 2, 302-309 (1988).
Patel, P. I., et al; Genetic Mapping of Autosomal Dominant Charcot-Marie-Tooth Diseases in a Large French-Acadian Kindred: Identification of New Linked Markers on Chromosome 17; Am. J. Hum. Genet., 46:802-809 (1990).
Patel, P. I., et al; Isolation of a Marker Linked to the Charcot-Marie-Tooth Disease Type IA Gene by Differential Alu-PCR of Human Chromosome 17-retaining Hybrids; Am. J. Hum. Genet., 47-926-934 (1990).
Raeymaekers, P., et al; Localization of the Mutation in an Extended Family with Charcot-Marie-Tooth Neuropathy (HMSN I); Am. J. Hum. Genet., 45:953-958 (1989).
Ray, R., et al; Three Polymorphisms at the D17S29 Locus; Nucleic Acids Research, 18:4958 (1990).
Schwartz, D. C. and Cantor, C. R.; Separation of Yeast Chromosome-Sized DNAs by Pulsed Field Gradient Gel Electrophoresis; Cell, 37:67-75 (1984).
Shyamala, V., et al; Tandem Chromosomal Duplications: Role of REP Sequences in the Recombination Event at the Join-Point. The EMBO Journal; 9:939-946 (1990).
Southern, E. M. Detection of Specific Sequences Among DNA Fragments Separated by Gel Electrophoresis; J. Mol. Biol., 98-503-517 (1975).
Sullivan,
This invention was made with government support under Grant No. NS-27042 awarded by the National Institute of Health. The government has certain rights in this invention.
Cardenas Odila S.
de Oca-Luna Roberto M.
Lupski James R.
Patel Pragna I.
Baylor College of Medicine
Bugaisky Gabriele E.
Wax Robert A.
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