Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1995-02-27
1997-11-18
Jones, W. Gary
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
935 77, 935 78, C12Q 168, C12N 1500
Patent
active
056886438
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
This invention relates to a method for differentiating a nucleic acid and an assay kit for such nucleic acid-differentiation. More specifically, this invention relates to a nucleic acid differentiating method that is capable of directly and quickly evaluating a specimen for the presence/absence of a mutant gene in a nucleic acid as well as the ratio of mutant to normal genes and that enables various assays that have been difficult to accomplish by conventional methods, for example, detection of a mutation at a non-specific site within a gene, detection of a small amount of abnormal cells containing a mutant gene in a mixture with normal cells, and determination of sequence matching of a particular gene in a plurality of samples; and an assay kit which may be used for nucleic acid-differentiation by the same method.
BACKGROUND ART
Molecular biology and genetics have experienced a marked progress in recent years, and the findings accumulated in such fields would contribute not only to chemical/physical clarification of various phyenomena involved in life, but also to human lives, in particular, to medicine as a field and a practice. DNA medicine starting from DNA is expanding to a clinical field far beyond expectation. It is known today that almost all diseases are related to DNA, and a diagnosis on a genetic level has become indispensable.
It is now recognized that nearly all enzyme deficiencies that are known for many years as inborn errors of metabolism are ascribed to diseases that are generally referred to as genetic diseases (molecular diseases). Detection of a mutation in a gene is quite effective for the diagnosis of such genetic diseases.
Typical conventional methods for detecting a mutation in a gene that can be used when the site of the genetic mutation is already known include a detection method using an oligonucleotide probe (PNAS, 80, 278, 1983): a method utilizing restriction enzyme polymorphism (Am. J. Hum. Genet., 69, 201, 1980); and scission of a one-base mismatch in a RNA:DNA hybrid by utilizing ribonuclease (Science, 230, 1243, 1985).
Mutation detection utilizing gene amplification was also developed (Proc. Natl. Acad. Sci. USA, 88, 189, 1991); Anal. Biochem, 186, 64-68, 1990). These methods, however, can be used only when the base sequence has already been found out and are limited to the detection of a specific mutation. On the other hand, SSCP method (Proc. Natl. Acad. Sci. USA, 86, 2766, 1989), DGGE method (Proc. Natl. Acad. Sci. USA, 86, 232, 1989), and other methods were reported as methods for detecting a non-specific mutation (site and base) in a certain region. These methods, however, use electrophoresis and are not necessarily practical when the ease and quickness of operation are taken into account.
Then J. C. Nicolas developed the following method as a system for detecting a non-specific mutation within a certain region of a nucleic acid (EP-A 362042 and Terouanne et al. Anal. Biochem., 205, 193, 1992). First a biotin label is introduced into one of duplex strands in a nucleic acid fragment containing a region to be detected for mutation, and an FITC label is introduced into the other strand to thereby produce a labeled standard DNA. The labeled standard DNA is mixed with an excessive amount of a sample DNA containing an unlabeled nucleic acid fragment of the same region as the standard DNA. The mixture is heated for denaturing and then slowly cooled down (competitive hybridization). When a fragment with a base sequence in complete conformity with that of the labeled standard DNA is present in the sample, recombination of double stranded DNA occurs between the duplex strands of the labeled standard DNA and the duplex strands of the sample DNA, whereupon the amount of the labeled standard DNA having both the biotin label and the FITC label that has been initially present is reduced. On the other hand, when a fragment with a base sequence partly different from that of the labeled standard DNA is present in the sample, the aforementioned recombination between
REFERENCES:
patent: 4605735 (1986-08-01), Miyoshi et al.
patent: 4667025 (1987-05-01), Miyoshi et al.
patent: 4789737 (1988-12-01), Miyoshi et al.
patent: 4849336 (1989-07-01), Miyoshi et al.
Conner et al, Proc. Natl. Acad. Sci. USA, vol. 80, pp. 278-282 (Jan. 1983).
Kohonen-Corish et al, Am. J. Human Genetics, vol. 39, pp. 751-762 (1986).
Myers et al, Science, vol. 230, pp. 1242-1246 (Dec. 13, 1985).
Sarkar et al, Analytical Biochemistry, vol. 186, pp. 64-68 (1990).
Terouanne et al, Analytical Biochemistry, vol. 205, pp. 193-199 (1992).
Barany, Proc. Natl. Acad. Sci. USA, vol. 88, pp. 189-193 (Jan. 1991).
Sheffield et al, Proc. Natl. Acad. Sci. USA, vol. 86, pp. 232-236 (Jan. 1989).
Orita et al, Proc. Natl. Acad. Sci. USA, vol. 86, pp. 2766-2770 (Apr. 1989).
Ubukata et al, Journal of Clinical Microbiology, vol. 30, No. 7, pp. 1728-1733 (Jul. 1992).
PCR Protocols, Academic Press Inc., pp. 14-15 and 352-353 (1990).
PCR Technology, Henry A. Erlich, Ed., Stockton Press (1989) (Introductory pages only).
Saiki et al, Nature, vol. 324, pp. 163-168 (Nov. 1986).
Maeda et al, Tissue Antigens, vol. 34, pp. 290-298 (1989).
Olerup et al, Tissue Antigens, vol. 39, pp. 225-235 (1992).
Jikken Igaku, Experimental Medicine, Yohdo-sha, vol. 8, No. 9 (1990).
Sekiya et al, Gann, vol. 74, pp. 794-797 (Dec. 1983).
Yuasa et al, Nature, vol. 303, pp. 775-779 (Jun. 30, 1983).
Ho et al, Gene, vol. 77, pp. 51-59 (1989).
The Stratagene Catalog p. 39 (1988).
Terouanne et al, Analytical Biochemistry. 205, 193-199 (1992).
Matsunaga Hironari
Oka Takanori
Yamane Akio
Jones W. Gary
Wakunaga Seiyaku Kabushiki Kaisha
Whisenant Ethan
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