Comparison of allele specific oligonucleotide-polymerase chain reaction and direct sequencing for high throughput screening of ABL kinase domain mutations in chronic myeloid leukemia resistant to imatinib.

Abstract	To identify a fast and sensitive method for screening for mutations in patients with imatinib- resistant chronic myeloid leukemia (CML), we compared allele specific oligonucleotide- polymerase chain reaction (ASO-PCR) assay with conventional direct sequencing. Among the 68 imatinib resistant CML patients studied, 18 amino acid substitutions were detected in 44 patients by two assays. The sensitivity of ASO-PCR was superior to that of direct sequencing as it could detect one mutant allele in 100 approximately 100,000 wild type sequences. The fastness, simplicity, and sensitivity of ASO-PCR assays will be useful for routine monitoring of mutations, especially for frequently identified mutations.
Authors	Ho-Young Kang, Ji-Yeon Hwang, Su-Hyun Kim, Hyun-gyung Goh, Myungshin Kim, Dong-Wook Kim
Journal	Haematologica (Haematologica) Vol. 91 Issue 5 Pg. 659-62 (May 2006) ISSN: 1592-8721 [Electronic] Italy
PMID	16627254 (Publication Type: Comparative Study, Evaluation Study, Journal Article, Research Support, Non-U.S. Gov't)
Chemical References	Antineoplastic Agents Benzamides Piperazines Protein Kinase Inhibitors Pyrimidines Imatinib Mesylate Fusion Proteins, bcr-abl
Topics	Alleles Amino Acid Substitution Antineoplastic Agents (pharmacology, therapeutic use) Benzamides Drug Resistance, Neoplasm (genetics) Fusion Proteins, bcr-abl (antagonists & inhibitors, genetics) Genes, abl Genetic Testing (methods) Humans Imatinib Mesylate K562 Cells (drug effects, enzymology) Leukemia, Myelogenous, Chronic, BCR-ABL Positive (drug therapy, enzymology, genetics) Mutation, Missense Piperazines (pharmacology, therapeutic use) Point Mutation Polymerase Chain Reaction (methods) Protein Kinase Inhibitors (pharmacology, therapeutic use) Pyrimidines (pharmacology, therapeutic use) Sensitivity and Specificity Sequence Analysis, DNA Time Factors

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