Enrichment of methylated molecules using enhanced-ice-co-amplification at lower denaturation temperature-PCR (E-ice-COLD-PCR) for the sensitive detection of disease-related hypermethylation
The detection of specific DNA methylation patterns bears great promise as biomarker for personalized management of cancer patients. Co-amplification at lower denaturation temperature-PCR (COLD-PCR) assays are sensitive methods, but have previously only been able to analyze loss of DNA methylation. E...
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| Veröffentlicht in: | Epigenomics 2018-05, Vol.10 (5), p.525-537 |
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| creator | Mauger, Florence Kernaleguen, Magali Lallemand, Céline Kristensen, Vessela N Deleuze, Jean-François Tost, Jörg |
| description | The detection of specific DNA methylation patterns bears great promise as biomarker for personalized management of cancer patients. Co-amplification at lower denaturation temperature-PCR (COLD-PCR) assays are sensitive methods, but have previously only been able to analyze loss of DNA methylation.
Enhanced (E)-ice-COLD-PCR reactions starting from 2 ng of bisulfite-converted DNA were developed to analyze methylation patterns in two promoters with locked nucleic acid (LNA) probes blocking amplification of unmethylated CpGs. The enrichment of methylated molecules was compared to quantitative (q)PCR and quantified using serial dilutions.
E-ice-COLD-PCR allowed the multiplexed enrichment and quantification of methylated DNA. Assays were validated in primary breast cancer specimens and circulating cell-free DNA from cancer patients.
E-ice-COLD-PCR could prove a useful tool in the context of DNA methylation analysis for personalized medicine. |
| doi_str_mv | 10.2217/epi-2017-0166 |
| format | Article |
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Enhanced (E)-ice-COLD-PCR reactions starting from 2 ng of bisulfite-converted DNA were developed to analyze methylation patterns in two promoters with locked nucleic acid (LNA) probes blocking amplification of unmethylated CpGs. The enrichment of methylated molecules was compared to quantitative (q)PCR and quantified using serial dilutions.
E-ice-COLD-PCR allowed the multiplexed enrichment and quantification of methylated DNA. Assays were validated in primary breast cancer specimens and circulating cell-free DNA from cancer patients.
E-ice-COLD-PCR could prove a useful tool in the context of DNA methylation analysis for personalized medicine.</description><identifier>ISSN: 1750-1911</identifier><identifier>EISSN: 1750-192X</identifier><identifier>DOI: 10.2217/epi-2017-0166</identifier><identifier>PMID: 29697281</identifier><language>eng</language><publisher>England: Future Medicine Ltd</publisher><subject>Amplification ; Biochemistry, Molecular Biology ; Biomarkers ; Bisulfite ; Breast cancer ; Cancer ; Cold ; Cold starts ; Denaturation ; DNA methylation ; DNA probes ; Enrichment ; Epigenetics ; Genetics ; Genomics ; Ice ; Life Sciences ; Mutation ; Nucleic acids ; Polymerase chain reaction ; Precision medicine</subject><ispartof>Epigenomics, 2018-05, Vol.10 (5), p.525-537</ispartof><rights>2018. This work is published under http://creativecommons.org/licenses/by-nc-nd/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>Attribution - NonCommercial - NoDerivatives</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c394t-b1d15b6822782e61d453102cdce07b3561e710a0df05111f2db499746a04fb713</citedby><cites>FETCH-LOGICAL-c394t-b1d15b6822782e61d453102cdce07b3561e710a0df05111f2db499746a04fb713</cites><orcidid>0000-0002-8402-4338</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29697281$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://cea.hal.science/cea-04485686$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Mauger, Florence</creatorcontrib><creatorcontrib>Kernaleguen, Magali</creatorcontrib><creatorcontrib>Lallemand, Céline</creatorcontrib><creatorcontrib>Kristensen, Vessela N</creatorcontrib><creatorcontrib>Deleuze, Jean-François</creatorcontrib><creatorcontrib>Tost, Jörg</creatorcontrib><title>Enrichment of methylated molecules using enhanced-ice-co-amplification at lower denaturation temperature-PCR (E-ice-COLD-PCR) for the sensitive detection of disease-related hypermethylation</title><title>Epigenomics</title><addtitle>Epigenomics</addtitle><description>The detection of specific DNA methylation patterns bears great promise as biomarker for personalized management of cancer patients. Co-amplification at lower denaturation temperature-PCR (COLD-PCR) assays are sensitive methods, but have previously only been able to analyze loss of DNA methylation.
Enhanced (E)-ice-COLD-PCR reactions starting from 2 ng of bisulfite-converted DNA were developed to analyze methylation patterns in two promoters with locked nucleic acid (LNA) probes blocking amplification of unmethylated CpGs. The enrichment of methylated molecules was compared to quantitative (q)PCR and quantified using serial dilutions.
E-ice-COLD-PCR allowed the multiplexed enrichment and quantification of methylated DNA. Assays were validated in primary breast cancer specimens and circulating cell-free DNA from cancer patients.
E-ice-COLD-PCR could prove a useful tool in the context of DNA methylation analysis for personalized medicine.</description><subject>Amplification</subject><subject>Biochemistry, Molecular Biology</subject><subject>Biomarkers</subject><subject>Bisulfite</subject><subject>Breast cancer</subject><subject>Cancer</subject><subject>Cold</subject><subject>Cold starts</subject><subject>Denaturation</subject><subject>DNA methylation</subject><subject>DNA probes</subject><subject>Enrichment</subject><subject>Epigenetics</subject><subject>Genetics</subject><subject>Genomics</subject><subject>Ice</subject><subject>Life Sciences</subject><subject>Mutation</subject><subject>Nucleic acids</subject><subject>Polymerase chain reaction</subject><subject>Precision medicine</subject><issn>1750-1911</issn><issn>1750-192X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><sourceid>BENPR</sourceid><recordid>eNpdksFu1DAQhiMEolXpkSuyxKU9GDxOYifHartQpJWKEEjcLMeZEFdJvNhO0b4NT8KBJ8Np2j3giz3WN7_-0fxZ9hrYO85Bvse9pZyBpAyEeJadgiwZhZp_f358A5xk5yHcsXRyXtUCXmYnvBa15BWcZn-2k7emH3GKxHVkxNgfBh2xJaMb0MwDBjIHO_0gOPV6MthSa5AaR_W4H2xnjY7WTURHMrhf6EmLk46zX38jjnv0S4308-YLudg-dG9ud9dLffn3d-c8iT2SgFOw0d5jEohoHrqTn9YG1AGpx9VUf0h6TyYT8yp70ekh4PnjfZZ9-7D9urmhu9uPnzZXO2ryuoi0gRbKRlScy4qjgLYoc2DctAaZbPJSAEpgmrUdKwGg421T1LUshGZF10jIz7LLVbfXg9p7O2p_UE5bdXO1Uwa1YkVRlaIS9wt7sbJ7737OGKIabTA4DHpCNwfFWQ4FFJKVCX37H3rnZj-lSVTaryhzLoAniq6U8S4Ej93RAbCFkyrlQC05UEsOEv_mUXVuRmyP9NPW839v27Af</recordid><startdate>20180501</startdate><enddate>20180501</enddate><creator>Mauger, Florence</creator><creator>Kernaleguen, Magali</creator><creator>Lallemand, Céline</creator><creator>Kristensen, Vessela N</creator><creator>Deleuze, Jean-François</creator><creator>Tost, Jörg</creator><general>Future Medicine Ltd</general><general>Future Medicine</general><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>EHMNL</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P5Z</scope><scope>P62</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>1XC</scope><scope>VOOES</scope><orcidid>https://orcid.org/0000-0002-8402-4338</orcidid></search><sort><creationdate>20180501</creationdate><title>Enrichment of methylated molecules using enhanced-ice-co-amplification at lower denaturation temperature-PCR (E-ice-COLD-PCR) for the sensitive detection of disease-related hypermethylation</title><author>Mauger, Florence ; 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Co-amplification at lower denaturation temperature-PCR (COLD-PCR) assays are sensitive methods, but have previously only been able to analyze loss of DNA methylation.
Enhanced (E)-ice-COLD-PCR reactions starting from 2 ng of bisulfite-converted DNA were developed to analyze methylation patterns in two promoters with locked nucleic acid (LNA) probes blocking amplification of unmethylated CpGs. The enrichment of methylated molecules was compared to quantitative (q)PCR and quantified using serial dilutions.
E-ice-COLD-PCR allowed the multiplexed enrichment and quantification of methylated DNA. Assays were validated in primary breast cancer specimens and circulating cell-free DNA from cancer patients.
E-ice-COLD-PCR could prove a useful tool in the context of DNA methylation analysis for personalized medicine.</abstract><cop>England</cop><pub>Future Medicine Ltd</pub><pmid>29697281</pmid><doi>10.2217/epi-2017-0166</doi><tpages>13</tpages><orcidid>https://orcid.org/0000-0002-8402-4338</orcidid><oa>free_for_read</oa></addata></record> |
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| source | PubMed Central |
| subjects | Amplification Biochemistry, Molecular Biology Biomarkers Bisulfite Breast cancer Cancer Cold Cold starts Denaturation DNA methylation DNA probes Enrichment Epigenetics Genetics Genomics Ice Life Sciences Mutation Nucleic acids Polymerase chain reaction Precision medicine |
| title | Enrichment of methylated molecules using enhanced-ice-co-amplification at lower denaturation temperature-PCR (E-ice-COLD-PCR) for the sensitive detection of disease-related hypermethylation |
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