Quantitative determination of the ratio of mutated to normal ras genes in the blood of leukemia patients by allele-specific PCR Journal Article
| Authors: | Horikoshi, T.; Lenz, H. J.; Danenberg, K.; Koch, O. M.; Bertino, J. R.; Danenberg, P. V. |
| Article Title: | Quantitative determination of the ratio of mutated to normal ras genes in the blood of leukemia patients by allele-specific PCR |
| Abstract: | By combining allele-specific PCR amplification and a PCR-based quantitation approach, a method has been developed to estimate the mutated K-ras gene content in the blood of AML patients as a percentage of total K-ras. One PCR primer set was designed not to discriminate between mutant K-ras and wild-type K-ras and thus amplified the total K-ras gene. The other PCR primer set was designed to be allele-specific for K-ras genes containing a G to C mutation at codon 12. This primer set could discriminate the mutant and wild-type genes when the proportion of the mutated sequence was 0.2% of the total K-ras gene. To test the method on biological specimens, genomic DNA samples were analyzed from the peripheral blood of a patient who had secondary AML with the same codon 12 K-ras mutation. Two samples taken from this patient 2 months apart during follow-up had myeloblast cell contents of 67 and 80%. However, the percentage of mutated K-ras was 50% in both samples, suggesting that this patient may be inherently heterozygotic in this particular mutation. This ratio of mutated to normal K-ras in the patient's cells was confirmed by RNA-SSCP analysis and RNA sequencing. This quantitation method can provide a sensitive and specific estimation of the content of mutated K-ras alleles in patient samples. © 1994. |
| Keywords: | controlled study; gene mutation; human cell; clinical trial; follow up; polymerase chain reaction; allele; controlled clinical trial; gene amplification; alleles; heterozygote; rna; dna; molecular sequence data; reference values; dna, neoplasm; acute myeloblastic leukemia; base sequence; neoplasms, second primary; dna primers; oncogene k ras; genes, ras; monocytes; codon; point mutation; dna determination; heterozygote detection; rna sequence; rna analysis; leukemia, myelocytic, acute; myeloblast; humans; human; priority journal; article; leukemia, mutation; quantitation, allele-specific pcr, ras |
| Journal Title: | Leukemia Research |
| Volume: | 18 |
| Issue: | 9 |
| ISSN: | 0145-2126 |
| Publisher: | Elsevier Ltd |
| Date Published: | 1994-09-01 |
| Start Page: | 693 |
| End Page: | 702 |
| Language: | English |
| DOI: | 10.1016/0145-2126(94)90069-8 |
| PROVIDER: | scopus |
| PUBMED: | 7934145 |
| DOI/URL: | |
| Notes: | Export Date: 14 January 2019 -- Article -- CODEN: LERED C2 -- Source: Scopus |
