Loading…
Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context
Purpose Translation of pharmacogenetic findings from the research laboratory to the clinical practice demands simple and efficient procedures. In this sense, we evaluated the suitability of a modified protocol for genomic DNA extraction based on alkaline lysis of cells. Methods Dried blood samples w...
Saved in:
| Published in: | Cancer chemotherapy and pharmacology 2015-05, Vol.75 (5), p.1095-1098 |
|---|---|
| Main Authors: | , , , , |
| Format: | Article |
| Language: | English |
| Subjects: | |
| Citations: | Items that this one cites Items that cite this one |
| Online Access: | Get full text |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| cited_by | cdi_FETCH-LOGICAL-c442t-d76aa36f6eb6ec49ac50f6ed6dff21f598fbccd40e885d6d4b1155503c2c49193 |
|---|---|
| cites | cdi_FETCH-LOGICAL-c442t-d76aa36f6eb6ec49ac50f6ed6dff21f598fbccd40e885d6d4b1155503c2c49193 |
| container_end_page | 1098 |
| container_issue | 5 |
| container_start_page | 1095 |
| container_title | Cancer chemotherapy and pharmacology |
| container_volume | 75 |
| creator | Ramos-Díaz, Ruth Gutiérrez-Nicolás, Fernando Nazco-Casariego, Gloria J González-Perera, Itamar Pérez-Pérez, José A |
| description | Purpose
Translation of pharmacogenetic findings from the research laboratory to the clinical practice demands simple and efficient procedures. In this sense, we evaluated the suitability of a modified protocol for genomic DNA extraction based on alkaline lysis of cells.
Methods
Dried blood samples were obtained from 48 patients diagnosed with colorectal cancer. A total of 11 mutations in the dihydropyrimidine dehydrogenase gene and related to 5-fluorouracil toxicity were searched by amplicon sequencing and real-time PCR with fluorescent probes.
Results
Genomic DNA extracted with the alkaline lysis method, both from dried blood samples and buccal swabs, fulfilled the quality requirements of the two genotyping methods assayed, which yielded 100 % concordant results for 11 genetic variants with relevance to cancer chemotherapy.
Conclusions
The assessed protocol has shown to be a very fast and economical approach to perform genetic analyses in the clinical laboratory for pharmacological purposes. |
| doi_str_mv | 10.1007/s00280-015-2729-4 |
| format | article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_journals_1678470765</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><sourcerecordid>3674080021</sourcerecordid><originalsourceid>FETCH-LOGICAL-c442t-d76aa36f6eb6ec49ac50f6ed6dff21f598fbccd40e885d6d4b1155503c2c49193</originalsourceid><addsrcrecordid>eNp1kMtOwzAQRS0EouXxAWyQJdaGsWPnsazKU6pgA2wtx7HblCQudiro3-OSgtiwGs3MmTPSReiMwiUFyK4CAMuBABWEZawgfA-NKU8YgZwn-2gMCedEZMBH6CiEJQBwmiSHaMREDjwyY7R4VU1dqb52HXYWK2xV6LHqKty4D6LdtmneItMZ3GxCHXBr-oWrsHUez68fJ9h89l7pb0HdRcFqoXyrtJubzvS1xtp1fWRO0IFVTTCnu3qMXm5vnqf3ZPZ09zCdzIjmnPWkylKlktSmpkyN5oXSAmJTpZW1jFpR5LbUuuJg8lzEKS8pFUJAolmkaZEco4vBu_LufW1CL5du7bv4UtI0y3kGWSoiRQdKexeCN1aufN0qv5EU5DZbOWQrY7Zym63k8eZ8Z16Xral-L37CjAAbgBBX3dz4P6__tX4BLmqFBQ</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1678470765</pqid></control><display><type>article</type><title>Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context</title><source>Springer Nature</source><creator>Ramos-Díaz, Ruth ; Gutiérrez-Nicolás, Fernando ; Nazco-Casariego, Gloria J ; González-Perera, Itamar ; Pérez-Pérez, José A</creator><creatorcontrib>Ramos-Díaz, Ruth ; Gutiérrez-Nicolás, Fernando ; Nazco-Casariego, Gloria J ; González-Perera, Itamar ; Pérez-Pérez, José A</creatorcontrib><description>Purpose
Translation of pharmacogenetic findings from the research laboratory to the clinical practice demands simple and efficient procedures. In this sense, we evaluated the suitability of a modified protocol for genomic DNA extraction based on alkaline lysis of cells.
Methods
Dried blood samples were obtained from 48 patients diagnosed with colorectal cancer. A total of 11 mutations in the dihydropyrimidine dehydrogenase gene and related to 5-fluorouracil toxicity were searched by amplicon sequencing and real-time PCR with fluorescent probes.
Results
Genomic DNA extracted with the alkaline lysis method, both from dried blood samples and buccal swabs, fulfilled the quality requirements of the two genotyping methods assayed, which yielded 100 % concordant results for 11 genetic variants with relevance to cancer chemotherapy.
Conclusions
The assessed protocol has shown to be a very fast and economical approach to perform genetic analyses in the clinical laboratory for pharmacological purposes.</description><identifier>ISSN: 0344-5704</identifier><identifier>EISSN: 1432-0843</identifier><identifier>DOI: 10.1007/s00280-015-2729-4</identifier><identifier>PMID: 25804843</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Alkalies - chemistry ; Cancer Research ; Colorectal Neoplasms - blood ; Colorectal Neoplasms - drug therapy ; Colorectal Neoplasms - genetics ; Dihydrouracil Dehydrogenase (NADP) - genetics ; DNA, Neoplasm - blood ; DNA, Neoplasm - genetics ; DNA, Neoplasm - isolation & purification ; Fluorouracil - toxicity ; Genotyping Techniques ; Humans ; Medicine ; Medicine & Public Health ; Mutation ; Oncology ; Pharmacogenetics ; Pharmacology/Toxicology ; Prospective Studies ; Reproducibility of Results ; Short Communication</subject><ispartof>Cancer chemotherapy and pharmacology, 2015-05, Vol.75 (5), p.1095-1098</ispartof><rights>Springer-Verlag Berlin Heidelberg 2015</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c442t-d76aa36f6eb6ec49ac50f6ed6dff21f598fbccd40e885d6d4b1155503c2c49193</citedby><cites>FETCH-LOGICAL-c442t-d76aa36f6eb6ec49ac50f6ed6dff21f598fbccd40e885d6d4b1155503c2c49193</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25804843$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ramos-Díaz, Ruth</creatorcontrib><creatorcontrib>Gutiérrez-Nicolás, Fernando</creatorcontrib><creatorcontrib>Nazco-Casariego, Gloria J</creatorcontrib><creatorcontrib>González-Perera, Itamar</creatorcontrib><creatorcontrib>Pérez-Pérez, José A</creatorcontrib><title>Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context</title><title>Cancer chemotherapy and pharmacology</title><addtitle>Cancer Chemother Pharmacol</addtitle><addtitle>Cancer Chemother Pharmacol</addtitle><description>Purpose
Translation of pharmacogenetic findings from the research laboratory to the clinical practice demands simple and efficient procedures. In this sense, we evaluated the suitability of a modified protocol for genomic DNA extraction based on alkaline lysis of cells.
Methods
Dried blood samples were obtained from 48 patients diagnosed with colorectal cancer. A total of 11 mutations in the dihydropyrimidine dehydrogenase gene and related to 5-fluorouracil toxicity were searched by amplicon sequencing and real-time PCR with fluorescent probes.
Results
Genomic DNA extracted with the alkaline lysis method, both from dried blood samples and buccal swabs, fulfilled the quality requirements of the two genotyping methods assayed, which yielded 100 % concordant results for 11 genetic variants with relevance to cancer chemotherapy.
Conclusions
The assessed protocol has shown to be a very fast and economical approach to perform genetic analyses in the clinical laboratory for pharmacological purposes.</description><subject>Alkalies - chemistry</subject><subject>Cancer Research</subject><subject>Colorectal Neoplasms - blood</subject><subject>Colorectal Neoplasms - drug therapy</subject><subject>Colorectal Neoplasms - genetics</subject><subject>Dihydrouracil Dehydrogenase (NADP) - genetics</subject><subject>DNA, Neoplasm - blood</subject><subject>DNA, Neoplasm - genetics</subject><subject>DNA, Neoplasm - isolation & purification</subject><subject>Fluorouracil - toxicity</subject><subject>Genotyping Techniques</subject><subject>Humans</subject><subject>Medicine</subject><subject>Medicine & Public Health</subject><subject>Mutation</subject><subject>Oncology</subject><subject>Pharmacogenetics</subject><subject>Pharmacology/Toxicology</subject><subject>Prospective Studies</subject><subject>Reproducibility of Results</subject><subject>Short Communication</subject><issn>0344-5704</issn><issn>1432-0843</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><recordid>eNp1kMtOwzAQRS0EouXxAWyQJdaGsWPnsazKU6pgA2wtx7HblCQudiro3-OSgtiwGs3MmTPSReiMwiUFyK4CAMuBABWEZawgfA-NKU8YgZwn-2gMCedEZMBH6CiEJQBwmiSHaMREDjwyY7R4VU1dqb52HXYWK2xV6LHqKty4D6LdtmneItMZ3GxCHXBr-oWrsHUez68fJ9h89l7pb0HdRcFqoXyrtJubzvS1xtp1fWRO0IFVTTCnu3qMXm5vnqf3ZPZ09zCdzIjmnPWkylKlktSmpkyN5oXSAmJTpZW1jFpR5LbUuuJg8lzEKS8pFUJAolmkaZEco4vBu_LufW1CL5du7bv4UtI0y3kGWSoiRQdKexeCN1aufN0qv5EU5DZbOWQrY7Zym63k8eZ8Z16Xral-L37CjAAbgBBX3dz4P6__tX4BLmqFBQ</recordid><startdate>20150501</startdate><enddate>20150501</enddate><creator>Ramos-Díaz, Ruth</creator><creator>Gutiérrez-Nicolás, Fernando</creator><creator>Nazco-Casariego, Gloria J</creator><creator>González-Perera, Itamar</creator><creator>Pérez-Pérez, José A</creator><general>Springer Berlin Heidelberg</general><general>Springer Nature B.V</general><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>7TO</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope></search><sort><creationdate>20150501</creationdate><title>Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context</title><author>Ramos-Díaz, Ruth ; Gutiérrez-Nicolás, Fernando ; Nazco-Casariego, Gloria J ; González-Perera, Itamar ; Pérez-Pérez, José A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c442t-d76aa36f6eb6ec49ac50f6ed6dff21f598fbccd40e885d6d4b1155503c2c49193</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Alkalies - chemistry</topic><topic>Cancer Research</topic><topic>Colorectal Neoplasms - blood</topic><topic>Colorectal Neoplasms - drug therapy</topic><topic>Colorectal Neoplasms - genetics</topic><topic>Dihydrouracil Dehydrogenase (NADP) - genetics</topic><topic>DNA, Neoplasm - blood</topic><topic>DNA, Neoplasm - genetics</topic><topic>DNA, Neoplasm - isolation & purification</topic><topic>Fluorouracil - toxicity</topic><topic>Genotyping Techniques</topic><topic>Humans</topic><topic>Medicine</topic><topic>Medicine & Public Health</topic><topic>Mutation</topic><topic>Oncology</topic><topic>Pharmacogenetics</topic><topic>Pharmacology/Toxicology</topic><topic>Prospective Studies</topic><topic>Reproducibility of Results</topic><topic>Short Communication</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ramos-Díaz, Ruth</creatorcontrib><creatorcontrib>Gutiérrez-Nicolás, Fernando</creatorcontrib><creatorcontrib>Nazco-Casariego, Gloria J</creatorcontrib><creatorcontrib>González-Perera, Itamar</creatorcontrib><creatorcontrib>Pérez-Pérez, José A</creatorcontrib><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>Oncogenes and Growth Factors Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Public Health Database</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</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 China</collection><jtitle>Cancer chemotherapy and pharmacology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ramos-Díaz, Ruth</au><au>Gutiérrez-Nicolás, Fernando</au><au>Nazco-Casariego, Gloria J</au><au>González-Perera, Itamar</au><au>Pérez-Pérez, José A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context</atitle><jtitle>Cancer chemotherapy and pharmacology</jtitle><stitle>Cancer Chemother Pharmacol</stitle><addtitle>Cancer Chemother Pharmacol</addtitle><date>2015-05-01</date><risdate>2015</risdate><volume>75</volume><issue>5</issue><spage>1095</spage><epage>1098</epage><pages>1095-1098</pages><issn>0344-5704</issn><eissn>1432-0843</eissn><abstract>Purpose
Translation of pharmacogenetic findings from the research laboratory to the clinical practice demands simple and efficient procedures. In this sense, we evaluated the suitability of a modified protocol for genomic DNA extraction based on alkaline lysis of cells.
Methods
Dried blood samples were obtained from 48 patients diagnosed with colorectal cancer. A total of 11 mutations in the dihydropyrimidine dehydrogenase gene and related to 5-fluorouracil toxicity were searched by amplicon sequencing and real-time PCR with fluorescent probes.
Results
Genomic DNA extracted with the alkaline lysis method, both from dried blood samples and buccal swabs, fulfilled the quality requirements of the two genotyping methods assayed, which yielded 100 % concordant results for 11 genetic variants with relevance to cancer chemotherapy.
Conclusions
The assessed protocol has shown to be a very fast and economical approach to perform genetic analyses in the clinical laboratory for pharmacological purposes.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>25804843</pmid><doi>10.1007/s00280-015-2729-4</doi><tpages>4</tpages></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0344-5704 |
| ispartof | Cancer chemotherapy and pharmacology, 2015-05, Vol.75 (5), p.1095-1098 |
| issn | 0344-5704 1432-0843 |
| language | eng |
| recordid | cdi_proquest_journals_1678470765 |
| source | Springer Nature |
| subjects | Alkalies - chemistry Cancer Research Colorectal Neoplasms - blood Colorectal Neoplasms - drug therapy Colorectal Neoplasms - genetics Dihydrouracil Dehydrogenase (NADP) - genetics DNA, Neoplasm - blood DNA, Neoplasm - genetics DNA, Neoplasm - isolation & purification Fluorouracil - toxicity Genotyping Techniques Humans Medicine Medicine & Public Health Mutation Oncology Pharmacogenetics Pharmacology/Toxicology Prospective Studies Reproducibility of Results Short Communication |
| title | Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context |
| url | http://sfxeu10.hosted.exlibrisgroup.com/loughborough?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-29T17%3A10%3A00IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_cross&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Validation%20of%20a%20fast%20and%20low-cost%20alkaline%20lysis%20method%20for%20gDNA%20extraction%20in%20a%20pharmacogenetic%20context&rft.jtitle=Cancer%20chemotherapy%20and%20pharmacology&rft.au=Ramos-D%C3%ADaz,%20Ruth&rft.date=2015-05-01&rft.volume=75&rft.issue=5&rft.spage=1095&rft.epage=1098&rft.pages=1095-1098&rft.issn=0344-5704&rft.eissn=1432-0843&rft_id=info:doi/10.1007/s00280-015-2729-4&rft_dat=%3Cproquest_cross%3E3674080021%3C/proquest_cross%3E%3Cgrp_id%3Ecdi_FETCH-LOGICAL-c442t-d76aa36f6eb6ec49ac50f6ed6dff21f598fbccd40e885d6d4b1155503c2c49193%3C/grp_id%3E%3Coa%3E%3C/oa%3E%3Curl%3E%3C/url%3E&rft_id=info:oai/&rft_pqid=1678470765&rft_id=info:pmid/25804843&rfr_iscdi=true |