Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1997-11-24
1999-09-07
Jones, W. Gary
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 231, 536 243, 935 76, 935 77, 935 78, C12Q 168, C07H 2102, C07H 2104, C12N 1500
Patent
active
059486180
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to a gene amplifying primer, a nucleic acid-differentiation method and a nucleic acid-differentiation assay kit using such primer. More specifically, the present invention relates to a gene amplifying primer which permits a target nucleic acid to be amplified, even when the target nucleic acid has a site of gene mutation or a difference among genes located in the vicinity of a primer binding site thereof, such that the site of gene mutation or difference may be shifted to the central side; a nucleic acid-differentiation method which ensures to detect a site of gene mutation or difference among genes in a target nucleic acid using the primer even when the site of gene mutation or difference is located near the primer binding site of the target nucleic acid; and a nucleic acid-differentiation assay kit for assaying genes by this nucleic acid-differentiation method.
BACKGROUND OF THE INVENTION
Recent years have brought amazing progress in the areas of molecular biology and genetics. The findings accumulated in such fields have not only contribution to the chemical and physical elucidation of biological phenomena, but also great influence on the human being, especially on medical science and practice. As a consequence, the DNA medicine, which started from DNA investigation, is marking great steps toward clinical fields far beyond the expectation. As it is recently revealed that almost all diseases are related to DNA, diagnosis at gene level is gradually regarded to be indispensable.
It is believed that those diseases now generally referred to as gene diseases (molecular diseases) include almost all of the enzyme deficiencies which are long known as diseases caused by inborn errors of metabolism. Detection of gene mutation is quite effective for the diagnosis of such gene diseases.
When a large number of genotypes are present as in the case of human leukocyte antigen (HLA), which hold the key to success of bone marrow transplantation or organ transplantation, it is desired to conveniently and rapidly determine the gene type at a high specificity.
An exemplary method for detecting the gene mutation or determining the gene type is the method of Nicolas, J. C. et al. (EP-A 362042; Anal. Biochem. 205, 193 (1992)), which is a modification of ED-PCR detection system (Japanese Patent Application Kokai (JP-A) Nos. 314965/1989 and 252300/1989; J. Clin. Microbiol. 30, 1728 (1992)).
The method of Nicolas et al. is carried out as follows. First, a labeled standard DNA is prepared from a double stranded nucleic acid fragment including a mutation region to be detected by introducing biotin label in one strand and FITC label in the other strand of the nucleic acid fragment. The labeled standard DNA is then mixed with an excess amount of a sample DNA including an unlabeled nucleic acid fragment of the same region as the standard DNA, and the mixture is heated for denaturation and slowly cooled (competitive hybridization). When a fragment including a nucleotide sequence identical with that of the labeled standard DNA is present in the sample, strand exchange would occur between the double stranded labeled standard DNA and the double stranded sample DNA, resulting in the amount of the labeled standard DNA possessing both the biotin label and the FITC label being reduced from the initial amount. On the other hand, when a fragment including a nucleotide sequence which is partly different from that of the labeled standard DNA is present in the sample, strand exchange is unlikely to occur between the double stranded labeled standard DNA and the double stranded sample DNA and the initial amount of the labeled standard DNA would remain substantially unchanged. Briefly stated, this method determines whether or not a fragment including a nucleotide sequence identical with that of the labeled standard DNA is present, by observing a change of the initial amount of the labeled standard DNA added through the steps of mixing, denaturation and annealing (competitive hybridization).
Also
REFERENCES:
patent: 5104792 (1992-04-01), Silver et al.
patent: 5106727 (1992-04-01), Hartley et al.
patent: 5629158 (1997-05-01), Uhlen
patent: 5688643 (1997-11-01), Oka et al.
Terouanne et al. (or Nicolas et al.), Analytical Biochemistry 205 : 193 (1992).
Oka et al., Nucleic Acids Research 22 (9) : 1541-1547 (1994).
Ubukata et al., J. of Clinical Microbiology 30 : 1728-1733 (1992).
Verrhasselt et al., DNA Sequence 2 :281-287 (1992).
Yamane et al., Nucleic Acids Research Symposium Series No. 20, pp. 91-92 (1988).
The Stratagene Catalog, p. 39 (1988 Edition).
Matthews et al., Analytical Biochemistry 169 : 1-25 (1988).
Innis et al., "PCR Protocols: A Guide to Methods and Application", pp. 84-91 and 909-112, Academic Press, San Diego, CA (1990).
Oka Takanori
Yamane Akio
Jones W. Gary
Wakunaga Seiyaku Kabushiki Kaisha
Whisenant Ethan
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