Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories
This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to...
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| Veröffentlicht in: | Mutagenesis 2014-07, Vol.29 (4), p.241-249 |
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| creator | Godschalk, Roger W L Ersson, Clara Stępnik, Maciej Ferlińska, Magdalena Palus, Jadwiga Teixeira, João Paulo Costa, Solange Jones, George D D Higgins, Jennifer A Kain, Johanna Möller, Lennart Forchhammer, Lykke Loft, Steffen Lorenzo, Yolanda Collins, Andrew R van Schooten, Frederik-Jan Laffon, Blanca Valdiglesias, Vanessa Cooke, Marcus Mistry, Vilas Karbaschi, Mahsa Phillips, David H Sozeri, Osman Routledge, Michael N Nelson-Smith, Kirsty Riso, Patrizia Porrini, Marisa López de Cerain, Adela Azqueta, Amaya Matullo, Giuseppe Allione, Alessandra Møller, Peter |
| description | This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp). |
| doi_str_mv | 10.1093/mutage/geu012 |
| format | Article |
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The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp).</description><identifier>ISSN: 0267-8357</identifier><identifier>EISSN: 1464-3804</identifier><identifier>DOI: 10.1093/mutage/geu012</identifier><identifier>PMID: 24737269</identifier><language>eng</language><publisher>England</publisher><subject>Adult ; Calibration ; Cell Separation - methods ; Comet Assay ; DNA Breaks, Double-Stranded ; DNA Damage ; DNA-Formamidopyrimidine Glycosylase - metabolism ; Female ; Humans ; Laboratories ; Leukocytes, Mononuclear - metabolism ; Middle Aged ; Mutagenicity Tests ; Regression Analysis</subject><ispartof>Mutagenesis, 2014-07, Vol.29 (4), p.241-249</ispartof><rights>The Author 2014. Published by Oxford University Press on behalf of the UK Environmental Mutagen Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c359t-98100f4258327fad4e102a1f9e5491dccb73848f88d99359a0253ca057694a663</citedby><cites>FETCH-LOGICAL-c359t-98100f4258327fad4e102a1f9e5491dccb73848f88d99359a0253ca057694a663</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,27901,27902</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24737269$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttp://kipublications.ki.se/Default.aspx?queryparsed=id:129272509$$DView record from Swedish Publication Index$$Hfree_for_read</backlink></links><search><creatorcontrib>Godschalk, Roger W L</creatorcontrib><creatorcontrib>Ersson, Clara</creatorcontrib><creatorcontrib>Stępnik, Maciej</creatorcontrib><creatorcontrib>Ferlińska, Magdalena</creatorcontrib><creatorcontrib>Palus, Jadwiga</creatorcontrib><creatorcontrib>Teixeira, João Paulo</creatorcontrib><creatorcontrib>Costa, Solange</creatorcontrib><creatorcontrib>Jones, George D D</creatorcontrib><creatorcontrib>Higgins, Jennifer A</creatorcontrib><creatorcontrib>Kain, Johanna</creatorcontrib><creatorcontrib>Möller, Lennart</creatorcontrib><creatorcontrib>Forchhammer, Lykke</creatorcontrib><creatorcontrib>Loft, Steffen</creatorcontrib><creatorcontrib>Lorenzo, Yolanda</creatorcontrib><creatorcontrib>Collins, Andrew R</creatorcontrib><creatorcontrib>van Schooten, Frederik-Jan</creatorcontrib><creatorcontrib>Laffon, Blanca</creatorcontrib><creatorcontrib>Valdiglesias, Vanessa</creatorcontrib><creatorcontrib>Cooke, Marcus</creatorcontrib><creatorcontrib>Mistry, Vilas</creatorcontrib><creatorcontrib>Karbaschi, Mahsa</creatorcontrib><creatorcontrib>Phillips, David H</creatorcontrib><creatorcontrib>Sozeri, Osman</creatorcontrib><creatorcontrib>Routledge, Michael N</creatorcontrib><creatorcontrib>Nelson-Smith, Kirsty</creatorcontrib><creatorcontrib>Riso, Patrizia</creatorcontrib><creatorcontrib>Porrini, Marisa</creatorcontrib><creatorcontrib>López de Cerain, Adela</creatorcontrib><creatorcontrib>Azqueta, Amaya</creatorcontrib><creatorcontrib>Matullo, Giuseppe</creatorcontrib><creatorcontrib>Allione, Alessandra</creatorcontrib><creatorcontrib>Møller, Peter</creatorcontrib><title>Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories</title><title>Mutagenesis</title><addtitle>Mutagenesis</addtitle><description>This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp).</description><subject>Adult</subject><subject>Calibration</subject><subject>Cell Separation - methods</subject><subject>Comet Assay</subject><subject>DNA Breaks, Double-Stranded</subject><subject>DNA Damage</subject><subject>DNA-Formamidopyrimidine Glycosylase - metabolism</subject><subject>Female</subject><subject>Humans</subject><subject>Laboratories</subject><subject>Leukocytes, Mononuclear - metabolism</subject><subject>Middle Aged</subject><subject>Mutagenicity Tests</subject><subject>Regression Analysis</subject><issn>0267-8357</issn><issn>1464-3804</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpNkMtOwzAQRS0EoqWwZIv8A6F-5eFlVZ5SBRtgG02ScQk4cWQnIP6eVCkVq7kanXsXh5BLzq4503LZDD1scbnFgXFxROZcJSqSGVPHZM5EkkaZjNMZOQvhgzGeioSdkplQqRyjnpPtG_ga-tq11Bl687SiFTTjILX4hTbQuqUd-rp7Rw-WFta5ijaude1QWgRPS7Q7KjgLPVY7vKqNQY9tTy0UzkPvfI3hnJwYsAEv9ndBXu9uX9YP0eb5_nG92kSljHUf6YwzZpSIMylSA5VCzgRwozFWmldlWaQyU5nJskrrsQFMxLIEFqeJVpAkckGiaTd8YzcUeefrBvxP7qDO96_PMWEeC8H1P770LgSP5tDgLN8JzifB-SR45K8mflxqsDrQf0blL06LejI</recordid><startdate>20140701</startdate><enddate>20140701</enddate><creator>Godschalk, Roger W L</creator><creator>Ersson, Clara</creator><creator>Stępnik, Maciej</creator><creator>Ferlińska, Magdalena</creator><creator>Palus, Jadwiga</creator><creator>Teixeira, João Paulo</creator><creator>Costa, Solange</creator><creator>Jones, George D D</creator><creator>Higgins, Jennifer A</creator><creator>Kain, Johanna</creator><creator>Möller, Lennart</creator><creator>Forchhammer, Lykke</creator><creator>Loft, Steffen</creator><creator>Lorenzo, Yolanda</creator><creator>Collins, Andrew R</creator><creator>van Schooten, Frederik-Jan</creator><creator>Laffon, Blanca</creator><creator>Valdiglesias, Vanessa</creator><creator>Cooke, Marcus</creator><creator>Mistry, Vilas</creator><creator>Karbaschi, Mahsa</creator><creator>Phillips, David H</creator><creator>Sozeri, Osman</creator><creator>Routledge, Michael N</creator><creator>Nelson-Smith, Kirsty</creator><creator>Riso, Patrizia</creator><creator>Porrini, Marisa</creator><creator>López de Cerain, Adela</creator><creator>Azqueta, Amaya</creator><creator>Matullo, Giuseppe</creator><creator>Allione, Alessandra</creator><creator>Møller, Peter</creator><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>ADTPV</scope><scope>AOWAS</scope></search><sort><creationdate>20140701</creationdate><title>Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories</title><author>Godschalk, Roger W L ; Ersson, Clara ; Stępnik, Maciej ; Ferlińska, Magdalena ; Palus, Jadwiga ; Teixeira, João Paulo ; Costa, Solange ; Jones, George D D ; Higgins, Jennifer A ; Kain, Johanna ; Möller, Lennart ; Forchhammer, Lykke ; Loft, Steffen ; Lorenzo, Yolanda ; Collins, Andrew R ; van Schooten, Frederik-Jan ; Laffon, Blanca ; Valdiglesias, Vanessa ; Cooke, Marcus ; Mistry, Vilas ; Karbaschi, Mahsa ; Phillips, David H ; Sozeri, Osman ; Routledge, Michael N ; Nelson-Smith, Kirsty ; Riso, Patrizia ; Porrini, Marisa ; López de Cerain, Adela ; Azqueta, Amaya ; Matullo, Giuseppe ; Allione, Alessandra ; Møller, Peter</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c359t-98100f4258327fad4e102a1f9e5491dccb73848f88d99359a0253ca057694a663</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Adult</topic><topic>Calibration</topic><topic>Cell Separation - methods</topic><topic>Comet Assay</topic><topic>DNA Breaks, Double-Stranded</topic><topic>DNA Damage</topic><topic>DNA-Formamidopyrimidine Glycosylase - metabolism</topic><topic>Female</topic><topic>Humans</topic><topic>Laboratories</topic><topic>Leukocytes, Mononuclear - metabolism</topic><topic>Middle Aged</topic><topic>Mutagenicity Tests</topic><topic>Regression Analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Godschalk, Roger W L</creatorcontrib><creatorcontrib>Ersson, Clara</creatorcontrib><creatorcontrib>Stępnik, Maciej</creatorcontrib><creatorcontrib>Ferlińska, Magdalena</creatorcontrib><creatorcontrib>Palus, Jadwiga</creatorcontrib><creatorcontrib>Teixeira, João Paulo</creatorcontrib><creatorcontrib>Costa, Solange</creatorcontrib><creatorcontrib>Jones, George D D</creatorcontrib><creatorcontrib>Higgins, Jennifer A</creatorcontrib><creatorcontrib>Kain, Johanna</creatorcontrib><creatorcontrib>Möller, Lennart</creatorcontrib><creatorcontrib>Forchhammer, Lykke</creatorcontrib><creatorcontrib>Loft, Steffen</creatorcontrib><creatorcontrib>Lorenzo, Yolanda</creatorcontrib><creatorcontrib>Collins, Andrew R</creatorcontrib><creatorcontrib>van Schooten, Frederik-Jan</creatorcontrib><creatorcontrib>Laffon, Blanca</creatorcontrib><creatorcontrib>Valdiglesias, Vanessa</creatorcontrib><creatorcontrib>Cooke, Marcus</creatorcontrib><creatorcontrib>Mistry, Vilas</creatorcontrib><creatorcontrib>Karbaschi, Mahsa</creatorcontrib><creatorcontrib>Phillips, David H</creatorcontrib><creatorcontrib>Sozeri, Osman</creatorcontrib><creatorcontrib>Routledge, Michael N</creatorcontrib><creatorcontrib>Nelson-Smith, Kirsty</creatorcontrib><creatorcontrib>Riso, Patrizia</creatorcontrib><creatorcontrib>Porrini, Marisa</creatorcontrib><creatorcontrib>López de Cerain, Adela</creatorcontrib><creatorcontrib>Azqueta, Amaya</creatorcontrib><creatorcontrib>Matullo, Giuseppe</creatorcontrib><creatorcontrib>Allione, Alessandra</creatorcontrib><creatorcontrib>Møller, Peter</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>SwePub</collection><collection>SwePub Articles</collection><jtitle>Mutagenesis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Godschalk, Roger W L</au><au>Ersson, Clara</au><au>Stępnik, Maciej</au><au>Ferlińska, Magdalena</au><au>Palus, Jadwiga</au><au>Teixeira, João Paulo</au><au>Costa, Solange</au><au>Jones, George D D</au><au>Higgins, Jennifer A</au><au>Kain, Johanna</au><au>Möller, Lennart</au><au>Forchhammer, Lykke</au><au>Loft, Steffen</au><au>Lorenzo, Yolanda</au><au>Collins, Andrew R</au><au>van Schooten, Frederik-Jan</au><au>Laffon, Blanca</au><au>Valdiglesias, Vanessa</au><au>Cooke, Marcus</au><au>Mistry, Vilas</au><au>Karbaschi, Mahsa</au><au>Phillips, David H</au><au>Sozeri, Osman</au><au>Routledge, Michael N</au><au>Nelson-Smith, Kirsty</au><au>Riso, Patrizia</au><au>Porrini, Marisa</au><au>López de Cerain, Adela</au><au>Azqueta, Amaya</au><au>Matullo, Giuseppe</au><au>Allione, Alessandra</au><au>Møller, Peter</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories</atitle><jtitle>Mutagenesis</jtitle><addtitle>Mutagenesis</addtitle><date>2014-07-01</date><risdate>2014</risdate><volume>29</volume><issue>4</issue><spage>241</spage><epage>249</epage><pages>241-249</pages><issn>0267-8357</issn><eissn>1464-3804</eissn><abstract>This study investigated the levels of DNA strand breaks and formamidopyrimidine DNA glycosylase (FPG) sensitive sites, as assessed by the comet assay, in peripheral blood mononuclear cells (PBMC) from healthy women from five different countries in Europe. The laboratory in each country (referred to as 'centre') collected and cryopreserved PBMC samples from three donors, using a standardised cell isolation protocol. The samples were analysed in 13 different laboratories for DNA damage, which is measured by the comet assay. The study aim was to assess variation in DNA damage in PBMC samples that were collected in the same way and processed using the same blood isolation procedure. The inter-laboratory variation was the prominent contributor to the overall variation. The inter-laboratory coefficient of variation decreased for both DNA strand breaks (from 68 to 26%) and FPG sensitive sites (from 57 to 12%) by standardisation of the primary comet assay endpoint with calibration curve samples. The level of DNA strand breaks in the samples from two of the centres (0.56-0.61 lesions/10(6) bp) was significantly higher compared with the other three centres (0.41-0.45 lesions/10(6) bp). In contrast, there was no difference between the levels of FPG sensitive sites in PBMC samples from healthy donors in the different centres (0.41-0.52 lesion/10(6) bp).</abstract><cop>England</cop><pmid>24737269</pmid><doi>10.1093/mutage/geu012</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0267-8357 |
| ispartof | Mutagenesis, 2014-07, Vol.29 (4), p.241-249 |
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| language | eng |
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| source | Oxford University Press Journals All Titles (1996-Current); MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Adult Calibration Cell Separation - methods Comet Assay DNA Breaks, Double-Stranded DNA Damage DNA-Formamidopyrimidine Glycosylase - metabolism Female Humans Laboratories Leukocytes, Mononuclear - metabolism Middle Aged Mutagenicity Tests Regression Analysis |
| title | Variation of DNA damage levels in peripheral blood mononuclear cells isolated in different laboratories |
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