Denaturing-HPLC-Based Assay for Detection of ABL Mutations in Chronic Myeloid Leukemia Patients Resistant to Imatinib
Despite the efficacy of the BCR-ABL tyrosine kinase inhibitor Imatinib mesylate for the treatment of chronic myeloid leukemia (CML), resistance has been observed in a proportion of cases, especially those with advanced stages of the disease. Point mutations within the ABL kinase domain are emerging...
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| Veröffentlicht in: | Clinical chemistry (Baltimore, Md.) Md.), 2004-07, Vol.50 (7), p.1205-1213 |
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| creator | Soverini, Simona Martinelli, Giovanni Amabile, Marilina Poerio, Angela Bianchini, Michele Rosti, Gianantonio Pane, Fabrizio Saglio, Giuseppe Baccarani, Michele |
| description | Despite the efficacy of the BCR-ABL tyrosine kinase inhibitor Imatinib mesylate for the treatment of chronic myeloid leukemia (CML), resistance has been observed in a proportion of cases, especially those with advanced stages of the disease. Point mutations within the ABL kinase domain are emerging as the most frequent mechanism for reactivation of kinase activity within the leukemic clone.
We developed a denaturing-HPLC (D-HPLC)-based assay for screening for ABL point mutations. For each sample, two partially overlapping fragments of 393 and 482 bp corresponding to the kinase domain were amplified by nested reverse transcription-PCR and analyzed under selected temperature and acetonitrile gradient conditions. Fifty-one bone marrow and/or peripheral blood specimens from 27 CML patients who showed cytogenetic resistance to Imatinib were screened in parallel by D-HPLC and by direct sequencing.
In 12 of 27 (44%) patients, D-HPLC showed an abnormal elution profile suggesting the presence of a nucleotide change. Direct sequencing confirmed the presence of a point mutation in all cases. Conversely, all samples scored as wild type by D-HPLC showed no evidence of mutations by direct sequencing. In two cases, novel amino acid substitutions at codons already known for being hot-spots of mutation were identified (F311I and E355D).
The proposed D-HPLC-based assay is highly specific and at least as sensitive as sequencing; with respect to the latter, it provides a much faster and less expensive semiautomated system for mutational screening. It may therefore potentially be a valuable tool for regular, large-scale testing of patients undergoing Imatinib treatment. |
| doi_str_mv | 10.1373/clinchem.2004.031112 |
| format | Article |
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We developed a denaturing-HPLC (D-HPLC)-based assay for screening for ABL point mutations. For each sample, two partially overlapping fragments of 393 and 482 bp corresponding to the kinase domain were amplified by nested reverse transcription-PCR and analyzed under selected temperature and acetonitrile gradient conditions. Fifty-one bone marrow and/or peripheral blood specimens from 27 CML patients who showed cytogenetic resistance to Imatinib were screened in parallel by D-HPLC and by direct sequencing.
In 12 of 27 (44%) patients, D-HPLC showed an abnormal elution profile suggesting the presence of a nucleotide change. Direct sequencing confirmed the presence of a point mutation in all cases. Conversely, all samples scored as wild type by D-HPLC showed no evidence of mutations by direct sequencing. In two cases, novel amino acid substitutions at codons already known for being hot-spots of mutation were identified (F311I and E355D).
The proposed D-HPLC-based assay is highly specific and at least as sensitive as sequencing; with respect to the latter, it provides a much faster and less expensive semiautomated system for mutational screening. It may therefore potentially be a valuable tool for regular, large-scale testing of patients undergoing Imatinib treatment.</description><identifier>ISSN: 0009-9147</identifier><identifier>EISSN: 1530-8561</identifier><identifier>DOI: 10.1373/clinchem.2004.031112</identifier><identifier>PMID: 15107311</identifier><identifier>CODEN: CLCHAU</identifier><language>eng</language><publisher>Washington, DC: Am Assoc Clin Chem</publisher><subject>Amino acids ; Analytical, structural and metabolic biochemistry ; Antineoplastic Agents - therapeutic use ; Benzamides ; Biological and medical sciences ; Bone marrow ; Chromatography, High Pressure Liquid - methods ; DNA Mutational Analysis ; Drug Resistance, Neoplasm ; Fundamental and applied biological sciences. Psychology ; Fusion Proteins, bcr-abl ; Genes, abl ; Hematologic and hematopoietic diseases ; Humans ; Imatinib Mesylate ; Investigative techniques, diagnostic techniques (general aspects) ; Leukemia ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - drug therapy ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics ; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis ; Liquid chromatography ; Medical sciences ; Molecular biophysics ; Mutation ; Piperazines - therapeutic use ; Point Mutation ; Protein-Tyrosine Kinases - genetics ; Pyrimidines - therapeutic use ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction ; Sensitivity and Specificity</subject><ispartof>Clinical chemistry (Baltimore, Md.), 2004-07, Vol.50 (7), p.1205-1213</ispartof><rights>2004 INIST-CNRS</rights><rights>Copyright American Association for Clinical Chemistry Jul 2004</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c457t-ca8e202a0508ce71f4e029f2b8051f249ebdd861ffaadbe775c7b0ac131a9cc93</citedby><cites>FETCH-LOGICAL-c457t-ca8e202a0508ce71f4e029f2b8051f249ebdd861ffaadbe775c7b0ac131a9cc93</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,778,782,27911,27912</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15918842$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/15107311$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Soverini, Simona</creatorcontrib><creatorcontrib>Martinelli, Giovanni</creatorcontrib><creatorcontrib>Amabile, Marilina</creatorcontrib><creatorcontrib>Poerio, Angela</creatorcontrib><creatorcontrib>Bianchini, Michele</creatorcontrib><creatorcontrib>Rosti, Gianantonio</creatorcontrib><creatorcontrib>Pane, Fabrizio</creatorcontrib><creatorcontrib>Saglio, Giuseppe</creatorcontrib><creatorcontrib>Baccarani, Michele</creatorcontrib><creatorcontrib>European LeukemiaNet-6th Framework Program of the European Community</creatorcontrib><creatorcontrib>Italian Cooerative Study Group on Chronic Myeloid Leukemia</creatorcontrib><creatorcontrib>the European LeukemiaNet—6th Framework Program of the European Community</creatorcontrib><creatorcontrib>the Italian Cooperative Study Group on Chronic Myeloid Leukemia (icsg-cml)</creatorcontrib><title>Denaturing-HPLC-Based Assay for Detection of ABL Mutations in Chronic Myeloid Leukemia Patients Resistant to Imatinib</title><title>Clinical chemistry (Baltimore, Md.)</title><addtitle>Clin Chem</addtitle><description>Despite the efficacy of the BCR-ABL tyrosine kinase inhibitor Imatinib mesylate for the treatment of chronic myeloid leukemia (CML), resistance has been observed in a proportion of cases, especially those with advanced stages of the disease. Point mutations within the ABL kinase domain are emerging as the most frequent mechanism for reactivation of kinase activity within the leukemic clone.
We developed a denaturing-HPLC (D-HPLC)-based assay for screening for ABL point mutations. For each sample, two partially overlapping fragments of 393 and 482 bp corresponding to the kinase domain were amplified by nested reverse transcription-PCR and analyzed under selected temperature and acetonitrile gradient conditions. Fifty-one bone marrow and/or peripheral blood specimens from 27 CML patients who showed cytogenetic resistance to Imatinib were screened in parallel by D-HPLC and by direct sequencing.
In 12 of 27 (44%) patients, D-HPLC showed an abnormal elution profile suggesting the presence of a nucleotide change. Direct sequencing confirmed the presence of a point mutation in all cases. Conversely, all samples scored as wild type by D-HPLC showed no evidence of mutations by direct sequencing. In two cases, novel amino acid substitutions at codons already known for being hot-spots of mutation were identified (F311I and E355D).
The proposed D-HPLC-based assay is highly specific and at least as sensitive as sequencing; with respect to the latter, it provides a much faster and less expensive semiautomated system for mutational screening. It may therefore potentially be a valuable tool for regular, large-scale testing of patients undergoing Imatinib treatment.</description><subject>Amino acids</subject><subject>Analytical, structural and metabolic biochemistry</subject><subject>Antineoplastic Agents - therapeutic use</subject><subject>Benzamides</subject><subject>Biological and medical sciences</subject><subject>Bone marrow</subject><subject>Chromatography, High Pressure Liquid - methods</subject><subject>DNA Mutational Analysis</subject><subject>Drug Resistance, Neoplasm</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Fusion Proteins, bcr-abl</subject><subject>Genes, abl</subject><subject>Hematologic and hematopoietic diseases</subject><subject>Humans</subject><subject>Imatinib Mesylate</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Leukemia</subject><subject>Leukemia, Myelogenous, Chronic, BCR-ABL Positive - drug therapy</subject><subject>Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics</subject><subject>Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis</subject><subject>Liquid chromatography</subject><subject>Medical sciences</subject><subject>Molecular biophysics</subject><subject>Mutation</subject><subject>Piperazines - therapeutic use</subject><subject>Point Mutation</subject><subject>Protein-Tyrosine Kinases - genetics</subject><subject>Pyrimidines - therapeutic use</subject><subject>Retrospective Studies</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Sensitivity and Specificity</subject><issn>0009-9147</issn><issn>1530-8561</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2004</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpdkdFqFDEUhoModlt9A5Eg6N2sSWaymVxut9YWplhEr0Mmc9JNnUlqkmHZtzfLrlTMTTjh-39O-BB6R8mS1qL-bEbnzRamJSOkWZKaUspeoAXlNalavqIv0YIQIitJG3GGzlN6LGMj2tVrdEY5JaIkFmi-Aq_zHJ1_qG7uu011qRMMeJ2S3mMbIr6CDCa74HGweH3Z4bs568OcsPN4s43BO4Pv9jAGN-AO5l8wOY3vCwM-J_wdkktZ-4xzwLdTefauf4NeWT0meHu6L9DP6y8_NjdV9-3r7WbdVabhIldGt8AI04ST1oCgtgHCpGV9Szi1rJHQD0O7otZqPfQgBDeiJ9rQmmppjKwv0Kdj71MMv2dIWU0uGRhH7SHMSa3KaVeSF_DDf-BjmKMvuylGaymFFE2BmiNkYkgpglVP0U067hUl6uBE_XWiDk7U0UmJvT91z_0Ew3PoJKEAH0-ATkaPNmpvXPqHk7RtG_b8m6172O5cBJUmPY6llqrdbseJEooywus_7MCkNQ</recordid><startdate>20040701</startdate><enddate>20040701</enddate><creator>Soverini, Simona</creator><creator>Martinelli, Giovanni</creator><creator>Amabile, Marilina</creator><creator>Poerio, Angela</creator><creator>Bianchini, Michele</creator><creator>Rosti, Gianantonio</creator><creator>Pane, Fabrizio</creator><creator>Saglio, Giuseppe</creator><creator>Baccarani, Michele</creator><general>Am Assoc Clin Chem</general><general>American Association for Clinical Chemistry</general><general>Oxford University Press</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>4U-</scope><scope>7QO</scope><scope>7RV</scope><scope>7TM</scope><scope>7U7</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>BKSAR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>NAPCQ</scope><scope>P64</scope><scope>PCBAR</scope><scope>PDBOC</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>Q9U</scope><scope>RC3</scope><scope>S0X</scope><scope>7X8</scope></search><sort><creationdate>20040701</creationdate><title>Denaturing-HPLC-Based Assay for Detection of ABL Mutations in Chronic Myeloid Leukemia Patients Resistant to Imatinib</title><author>Soverini, Simona ; Martinelli, Giovanni ; Amabile, Marilina ; Poerio, Angela ; Bianchini, Michele ; Rosti, Gianantonio ; Pane, Fabrizio ; Saglio, Giuseppe ; Baccarani, Michele</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c457t-ca8e202a0508ce71f4e029f2b8051f249ebdd861ffaadbe775c7b0ac131a9cc93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2004</creationdate><topic>Amino acids</topic><topic>Analytical, structural and metabolic biochemistry</topic><topic>Antineoplastic Agents - therapeutic use</topic><topic>Benzamides</topic><topic>Biological and medical sciences</topic><topic>Bone marrow</topic><topic>Chromatography, High Pressure Liquid - methods</topic><topic>DNA Mutational Analysis</topic><topic>Drug Resistance, Neoplasm</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Fusion Proteins, bcr-abl</topic><topic>Genes, abl</topic><topic>Hematologic and hematopoietic diseases</topic><topic>Humans</topic><topic>Imatinib Mesylate</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Leukemia</topic><topic>Leukemia, Myelogenous, Chronic, BCR-ABL Positive - drug therapy</topic><topic>Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics</topic><topic>Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis</topic><topic>Liquid chromatography</topic><topic>Medical sciences</topic><topic>Molecular biophysics</topic><topic>Mutation</topic><topic>Piperazines - therapeutic use</topic><topic>Point Mutation</topic><topic>Protein-Tyrosine Kinases - genetics</topic><topic>Pyrimidines - therapeutic use</topic><topic>Retrospective Studies</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Soverini, Simona</creatorcontrib><creatorcontrib>Martinelli, Giovanni</creatorcontrib><creatorcontrib>Amabile, Marilina</creatorcontrib><creatorcontrib>Poerio, Angela</creatorcontrib><creatorcontrib>Bianchini, Michele</creatorcontrib><creatorcontrib>Rosti, Gianantonio</creatorcontrib><creatorcontrib>Pane, Fabrizio</creatorcontrib><creatorcontrib>Saglio, Giuseppe</creatorcontrib><creatorcontrib>Baccarani, Michele</creatorcontrib><creatorcontrib>European LeukemiaNet-6th Framework Program of the European Community</creatorcontrib><creatorcontrib>Italian Cooerative Study Group on Chronic Myeloid Leukemia</creatorcontrib><creatorcontrib>the European LeukemiaNet—6th Framework Program of the European Community</creatorcontrib><creatorcontrib>the Italian Cooperative Study Group on Chronic Myeloid Leukemia (icsg-cml)</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>University Readers</collection><collection>Biotechnology Research Abstracts</collection><collection>Nursing & Allied Health Database</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Science Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Materials Science Collection</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Materials Science Database</collection><collection>Nursing & Allied Health Database (Alumni Edition)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Science Database</collection><collection>Nursing & Allied Health Premium</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Earth, Atmospheric & Aquatic Science Database</collection><collection>Materials Science Collection</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central Basic</collection><collection>Genetics Abstracts</collection><collection>SIRS Editorial</collection><collection>MEDLINE - Academic</collection><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Soverini, Simona</au><au>Martinelli, Giovanni</au><au>Amabile, Marilina</au><au>Poerio, Angela</au><au>Bianchini, Michele</au><au>Rosti, Gianantonio</au><au>Pane, Fabrizio</au><au>Saglio, Giuseppe</au><au>Baccarani, Michele</au><aucorp>European LeukemiaNet-6th Framework Program of the European Community</aucorp><aucorp>Italian Cooerative Study Group on Chronic Myeloid Leukemia</aucorp><aucorp>the European LeukemiaNet—6th Framework Program of the European Community</aucorp><aucorp>the Italian Cooperative Study Group on Chronic Myeloid Leukemia (icsg-cml)</aucorp><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Denaturing-HPLC-Based Assay for Detection of ABL Mutations in Chronic Myeloid Leukemia Patients Resistant to Imatinib</atitle><jtitle>Clinical chemistry (Baltimore, Md.)</jtitle><addtitle>Clin Chem</addtitle><date>2004-07-01</date><risdate>2004</risdate><volume>50</volume><issue>7</issue><spage>1205</spage><epage>1213</epage><pages>1205-1213</pages><issn>0009-9147</issn><eissn>1530-8561</eissn><coden>CLCHAU</coden><abstract>Despite the efficacy of the BCR-ABL tyrosine kinase inhibitor Imatinib mesylate for the treatment of chronic myeloid leukemia (CML), resistance has been observed in a proportion of cases, especially those with advanced stages of the disease. Point mutations within the ABL kinase domain are emerging as the most frequent mechanism for reactivation of kinase activity within the leukemic clone.
We developed a denaturing-HPLC (D-HPLC)-based assay for screening for ABL point mutations. For each sample, two partially overlapping fragments of 393 and 482 bp corresponding to the kinase domain were amplified by nested reverse transcription-PCR and analyzed under selected temperature and acetonitrile gradient conditions. Fifty-one bone marrow and/or peripheral blood specimens from 27 CML patients who showed cytogenetic resistance to Imatinib were screened in parallel by D-HPLC and by direct sequencing.
In 12 of 27 (44%) patients, D-HPLC showed an abnormal elution profile suggesting the presence of a nucleotide change. Direct sequencing confirmed the presence of a point mutation in all cases. Conversely, all samples scored as wild type by D-HPLC showed no evidence of mutations by direct sequencing. In two cases, novel amino acid substitutions at codons already known for being hot-spots of mutation were identified (F311I and E355D).
The proposed D-HPLC-based assay is highly specific and at least as sensitive as sequencing; with respect to the latter, it provides a much faster and less expensive semiautomated system for mutational screening. It may therefore potentially be a valuable tool for regular, large-scale testing of patients undergoing Imatinib treatment.</abstract><cop>Washington, DC</cop><pub>Am Assoc Clin Chem</pub><pmid>15107311</pmid><doi>10.1373/clinchem.2004.031112</doi><tpages>9</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0009-9147 |
| ispartof | Clinical chemistry (Baltimore, Md.), 2004-07, Vol.50 (7), p.1205-1213 |
| issn | 0009-9147 1530-8561 |
| language | eng |
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| source | MEDLINE; Oxford University Press Journals All Titles (1996-Current) |
| subjects | Amino acids Analytical, structural and metabolic biochemistry Antineoplastic Agents - therapeutic use Benzamides Biological and medical sciences Bone marrow Chromatography, High Pressure Liquid - methods DNA Mutational Analysis Drug Resistance, Neoplasm Fundamental and applied biological sciences. Psychology Fusion Proteins, bcr-abl Genes, abl Hematologic and hematopoietic diseases Humans Imatinib Mesylate Investigative techniques, diagnostic techniques (general aspects) Leukemia Leukemia, Myelogenous, Chronic, BCR-ABL Positive - drug therapy Leukemia, Myelogenous, Chronic, BCR-ABL Positive - genetics Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis Liquid chromatography Medical sciences Molecular biophysics Mutation Piperazines - therapeutic use Point Mutation Protein-Tyrosine Kinases - genetics Pyrimidines - therapeutic use Retrospective Studies Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity |
| title | Denaturing-HPLC-Based Assay for Detection of ABL Mutations in Chronic Myeloid Leukemia Patients Resistant to Imatinib |
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