Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1994-11-15
1996-04-30
Jones, W. Gary
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 18, 435 911, 435 912, 435 9152, 536 2433, C07H 2102, C07H 2104, C12P 1934, C12Q 168
Patent
active
055124410
ABSTRACT:
There is disclosed a quantitative sensitive method to enable the detection of point mutations at a known site to a diagnostic kit which uses a multi step (for example, four steps) or a single step reaction. The method uses selective polymerase chain reaction (PCR) amplification of mutant test gene sequences involving first stage amplification of both mutant and wild-type sequences, first stage restriction enzyme digestion of only wild-type sequences, second stage amplification of undigested amplified fragments enriched in mutant sequences and second stage digestion of previously undigested wild-type sequences. Long and short tail primers are used in the first and second stages of amplification respectively to enable selective amplification (in the second stage) of only previously amplified material and none of the original test genomic DNA. The short tail primers are labelled with biotin and fluorescence at their respective 5' and 3' ends to enable easy detection and quantitation of mutations in the test gene via automated fluorescence readers. The use of multi steps as well as a single step reaction is disclosed. The process is exemplified with respect to its use in detecting mutations in the human K-ras gene, yet it is applicable for any given mutation in a defined site.
REFERENCES:
patent: 4683195 (1987-07-01), Mullis et al.
patent: 4683202 (1987-07-01), Mullis et al.
patent: 4800159 (1989-01-01), Mullis et al.
patent: 4888278 (1989-12-01), Singer et al.
patent: 4889818 (1989-12-01), Gelfand et al.
patent: 4965188 (1990-10-01), Mullis et al.
patent: 4996142 (1991-02-01), Al-Hakim et al.
patent: 5024934 (1991-06-01), Lee
patent: 5049490 (1991-09-01), Sutherland et al.
patent: 5079352 (1992-01-01), Gelfand et al.
patent: 5187084 (1993-02-01), Hallisby
patent: 5210015 (1993-05-01), Gelfand et al.
patent: 5215899 (1993-06-01), Dattagupta
patent: 5225326 (1993-07-01), Bresser et al.
Innis et al., "PCR Protocols, A Guide to Methods and Applications," Academic Press, Inc., pp. 28-38, 92-98, 1990.
Detection of K-ras Mutation in Normal and Malignant Colonic Tissues by an Enriched PCR Method. Z. Ronai, et al Int'l Journal Oncology 4:391-6; 1994.
Rapid and Sensitive Non Radioactive Detection of Mutant K-ras Genes via Enriched PCR Amplification Scott Kahn, et al Oncogene (1991) 6, 1079-83.
Detection of Ki-ras Mutation in Non-Neoplastic Mucosa of Japanese Patients with Colorectal Cancers T. Minamoto, et al Int'l Journal Oncology 4:397-401 (1994).
Detection of K-ras Mutation in Colonic Effluent Samples from Patients Without Evidence of Colorectal Carcinoma M. Tobi, et al Journal of Nat. Cancer Inst. vol. 86 No. 13 1994 pp. 1007-1010.
The 1065 Air Thermo-Cycler User's Guide from Idaho Technology, pp. 1-5.
American Health Foundation
Jones W. Gary
Tran Paul B.
LandOfFree
Quantative method for early detection of mutant alleles and diag does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Quantative method for early detection of mutant alleles and diag, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Quantative method for early detection of mutant alleles and diag will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-627760