AustralieFrV1, Main, Exploration, bibRecord, 008417

Concordance of assays designed for the quantification of JAK2V617F : a multicenter study

Identifieur interne : 008417 ( Main/Exploration ); précédent : 008416; suivant : 008418

Concordance of assays designed for the quantification of JAK2V617F : a multicenter study

Auteurs : Eric Lippert [France] ; Francois Girodon [France] ; Emma Hammond [Australie] ; Jaroslav Jelinek [États-Unis] ; N. Scott Reading [États-Unis] ; Boris Fehse [Allemagne] ; Katy Hanlon [Royaume-Uni] ; Mirjam Hermans [Pays-Bas] ; Céline Richard [France] ; Sabina Swierczek [États-Unis] ; Valérie Ugo [France] ; Serge Carillo [France] ; Véronique Harrivel [France] ; Christophe Marzac [France] ; Daniela Pietra [Italie] ; Marta Sobas [Suisse] ; Morgane Mounier [France] ; Marina Migeon [France] ; Sian Ellard [Royaume-Uni] ; Nicolaus Kröger [Allemagne] ; Richard Herrmann [Australie] ; Josef T. Prchal [États-Unis] ; Radek C. Skoda [Suisse] ; Sylvie Hermouet [France]

Source :

Haematologica : (Roma) [ 0390-6078 ] ; 2009.

RBID : Pascal:09-0108409

Descripteurs français

Pascal (Inist)
- Réaction chaîne polymérase, Concordance, Analyse quantitative, Syndrome myéloprolifératif, Etude multicentrique, Normalisation, Allèle, Génétique, Hématologie, JAK2 protein tyrosine kinase.
Wicri :
- topic : Analyse quantitative, Normalisation, Génétique.

English descriptors

KwdEn :
- Allele, Concordance, Genetics, Hematology, JAK2 protein tyrosine kinase, Multicenter study, Myeloproliferative syndrome, Polymerase chain reaction, Quantitative analysis, Standardization.

Abstract

Background Many different techniques have been designed for the quantification of JAK2V617F allelic burden, sometimes producing discrepant results. Design and Methods JAK2V617F quantification techniques were compared among 16 centers using 11 assays based on quantitative polymerase chain reaction (with mutation-specific primers or probes, or fluorescent resonance energy transfer/melting curve analysis), allele-specific polymerase chain reaction, conventional sequencing or pyrosequencing. Results A first series of blinded samples (granulocyte DNA, n=29) was analyzed. Seven assays (12 centers) reported values inside the mean±2SD; the mean coefficient of variation was 31%. Sequencing techniques lacked sensitivity, and strong discrepancies were observed with four techniques, which could be attributed to inadequate standards or to different modes of expression of results. Indeed, quantification of JAK2V617F in relation to another control gene produced higher than expected values, suggesting the possibility of more than two JAK2 copies/cell. After calibration of assays with common 1% to 100% JAK2V617F standards (dilutions of UKE-1 cells in normal leukocytes), 14 centers tested ten new samples. JAK2V617F allelic burdens greater or equal than 1% were then reliably quantified by five techniques - one allele specific-polymerase chain reaction and four TaqMan allele-specific quantitative polymerase chain reaction assays, including one previously giving results outside the mean±2SD - with a lower mean coefficient of variation (21%). Of these, only the two TaqMan allele-specific quantitative polymerase chain reaction assays with primer-based specificity could detect 0.2% JAK2V617F. Conclusions Techniques expressing the allelic burden as JAK2V617F/total JAK2 and using a common set of standards produced similar quantification results but with variable sensitivity. Calibration to a reference standard improved reproducibility.

Affiliations:

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Le document en format XML

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<affiliation wicri:level="1"><inist:fA14 i1="08"><s1>Molecular Diagnostics, Jeroen Bosch Hospital</s1>
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<country>France</country>
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</placeName>
</affiliation>
</author>
<author><name sortKey="Swierczek, Sabina" sort="Swierczek, Sabina" uniqKey="Swierczek S" first="Sabina" last="Swierczek">Sabina Swierczek</name>
<affiliation wicri:level="2"><inist:fA14 i1="10"><s1>Hematology Division, University of Utah School of Medicine</s1>
<s2>Salt Lake City, Utah</s2>
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<sZ>10 aut.</sZ>
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<placeName><region type="state">Utah</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Ugo, Valerie" sort="Ugo, Valerie" uniqKey="Ugo V" first="Valérie" last="Ugo">Valérie Ugo</name>
<affiliation wicri:level="3"><inist:fA14 i1="11"><s1>Laboratoire d'Hématologie, CHU</s1>
<s2>Brest</s2>
<s3>FRA</s3>
<sZ>11 aut.</sZ>
</inist:fA14>
<country>France</country>
<placeName><region type="region">Région Bretagne</region>
<region type="old region">Région Bretagne</region>
<settlement type="city">Brest</settlement>
</placeName>
</affiliation>
</author>
<author><name sortKey="Carillo, Serge" sort="Carillo, Serge" uniqKey="Carillo S" first="Serge" last="Carillo">Serge Carillo</name>
<affiliation wicri:level="3"><inist:fA14 i1="12"><s1>Laboratoire de Cytologie Clinique et Cytogénétique, CHU</s1>
<s2>Nîmes</s2>
<s3>FRA</s3>
<sZ>12 aut.</sZ>
</inist:fA14>
<country>France</country>
<placeName><region type="region">Occitanie (région administrative)</region>
<region type="old region">Languedoc-Roussillon</region>
<settlement type="city">Nîmes</settlement>
</placeName>
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</author>
<author><name sortKey="Harrivel, Veronique" sort="Harrivel, Veronique" uniqKey="Harrivel V" first="Véronique" last="Harrivel">Véronique Harrivel</name>
<affiliation wicri:level="3"><inist:fA14 i1="13"><s1>Laboratoire d'Hématologie, Hôtel-Dieu</s1>
<s2>Paris</s2>
<s3>FRA</s3>
<sZ>13 aut.</sZ>
<sZ>14 aut.</sZ>
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<country>France</country>
<placeName><region type="region">Île-de-France</region>
<region type="old region">Île-de-France</region>
<settlement type="city">Paris</settlement>
</placeName>
</affiliation>
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<author><name sortKey="Marzac, Christophe" sort="Marzac, Christophe" uniqKey="Marzac C" first="Christophe" last="Marzac">Christophe Marzac</name>
<affiliation wicri:level="3"><inist:fA14 i1="13"><s1>Laboratoire d'Hématologie, Hôtel-Dieu</s1>
<s2>Paris</s2>
<s3>FRA</s3>
<sZ>13 aut.</sZ>
<sZ>14 aut.</sZ>
</inist:fA14>
<country>France</country>
<placeName><region type="region">Île-de-France</region>
<region type="old region">Île-de-France</region>
<settlement type="city">Paris</settlement>
</placeName>
</affiliation>
</author>
<author><name sortKey="Pietra, Daniela" sort="Pietra, Daniela" uniqKey="Pietra D" first="Daniela" last="Pietra">Daniela Pietra</name>
<affiliation wicri:level="1"><inist:fA14 i1="14"><s1>Department of Hematology, University of Pavia and Fondazione IRCCS Policlinico San Matteo</s1>
<s2>Pavia</s2>
<s3>ITA</s3>
<sZ>15 aut.</sZ>
</inist:fA14>
<country>Italie</country>
<wicri:noRegion>Pavia</wicri:noRegion>
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</author>
<author><name sortKey="Sobas, Marta" sort="Sobas, Marta" uniqKey="Sobas M" first="Marta" last="Sobas">Marta Sobas</name>
<affiliation wicri:level="1"><inist:fA14 i1="15"><s1>Experimental Hematology, Department of Biomedicine, Basel University Hospital</s1>
<s2>Basel</s2>
<s3>CHE</s3>
<sZ>16 aut.</sZ>
<sZ>23 aut.</sZ>
</inist:fA14>
<country>Suisse</country>
<wicri:noRegion>Basel</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Mounier, Morgane" sort="Mounier, Morgane" uniqKey="Mounier M" first="Morgane" last="Mounier">Morgane Mounier</name>
<affiliation wicri:level="3"><inist:fA14 i1="02"><s1>Laboratoire d'Hématologie, CHU</s1>
<s2>Dijon</s2>
<s3>FRA</s3>
<sZ>2 aut.</sZ>
<sZ>17 aut.</sZ>
</inist:fA14>
<country>France</country>
<placeName><region type="region">Bourgogne-Franche-Comté</region>
<region type="old region">Bourgogne</region>
<settlement type="city">Dijon</settlement>
</placeName>
</affiliation>
</author>
<author><name sortKey="Migeon, Marina" sort="Migeon, Marina" uniqKey="Migeon M" first="Marina" last="Migeon">Marina Migeon</name>
<affiliation wicri:level="3"><inist:fA14 i1="01"><s1>Laboratoire d'Hématologie, CHU</s1>
<s2>Bordeaux</s2>
<s3>FRA</s3>
<sZ>1 aut.</sZ>
<sZ>18 aut.</sZ>
</inist:fA14>
<country>France</country>
<placeName><region type="region">Nouvelle-Aquitaine</region>
<region type="old region">Aquitaine</region>
<settlement type="city">Bordeaux</settlement>
</placeName>
</affiliation>
</author>
<author><name sortKey="Ellard, Sian" sort="Ellard, Sian" uniqKey="Ellard S" first="Sian" last="Ellard">Sian Ellard</name>
<affiliation wicri:level="1"><inist:fA14 i1="07"><s1>Molecular Genetics, Royal Devon and Exeter Hospital</s1>
<s2>Exeter</s2>
<s3>GBR</s3>
<sZ>7 aut.</sZ>
<sZ>19 aut.</sZ>
</inist:fA14>
<country>Royaume-Uni</country>
<wicri:noRegion>Exeter</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Kroger, Nicolaus" sort="Kroger, Nicolaus" uniqKey="Kroger N" first="Nicolaus" last="Kröger">Nicolaus Kröger</name>
<affiliation wicri:level="3"><inist:fA14 i1="06"><s1>Uniklinik Eppendorf</s1>
<s2>Hamburg</s2>
<s3>DEU</s3>
<sZ>6 aut.</sZ>
<sZ>20 aut.</sZ>
</inist:fA14>
<country>Allemagne</country>
<placeName><settlement type="city">Hambourg</settlement>
<region type="land" nuts="2">Hambourg</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Herrmann, Richard" sort="Herrmann, Richard" uniqKey="Herrmann R" first="Richard" last="Herrmann">Richard Herrmann</name>
<affiliation wicri:level="1"><inist:fA14 i1="03"><s1>Murdoch University</s1>
<s2>Perth</s2>
<s3>AUS</s3>
<sZ>3 aut.</sZ>
<sZ>21 aut.</sZ>
</inist:fA14>
<country>Australie</country>
<wicri:noRegion>Murdoch University</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Prchal, Josef T" sort="Prchal, Josef T" uniqKey="Prchal J" first="Josef T." last="Prchal">Josef T. Prchal</name>
<affiliation wicri:level="2"><inist:fA14 i1="05"><s1>ARUP Institute for Clinical and Experimental Pathology</s1>
<s2>Salt Lake City, Utah</s2>
<s3>USA</s3>
<sZ>5 aut.</sZ>
<sZ>22 aut.</sZ>
</inist:fA14>
<country>États-Unis</country>
<placeName><region type="state">Utah</region>
</placeName>
</affiliation>
<affiliation wicri:level="2"><inist:fA14 i1="10"><s1>Hematology Division, University of Utah School of Medicine</s1>
<s2>Salt Lake City, Utah</s2>
<s3>USA</s3>
<sZ>10 aut.</sZ>
<sZ>22 aut.</sZ>
</inist:fA14>
<country>États-Unis</country>
<placeName><region type="state">Utah</region>
</placeName>
</affiliation>
</author>
<author><name sortKey="Skoda, Radek C" sort="Skoda, Radek C" uniqKey="Skoda R" first="Radek C." last="Skoda">Radek C. Skoda</name>
<affiliation wicri:level="1"><inist:fA14 i1="15"><s1>Experimental Hematology, Department of Biomedicine, Basel University Hospital</s1>
<s2>Basel</s2>
<s3>CHE</s3>
<sZ>16 aut.</sZ>
<sZ>23 aut.</sZ>
</inist:fA14>
<country>Suisse</country>
<wicri:noRegion>Basel</wicri:noRegion>
</affiliation>
</author>
<author><name sortKey="Hermouet, Sylvie" sort="Hermouet, Sylvie" uniqKey="Hermouet S" first="Sylvie" last="Hermouet">Sylvie Hermouet</name>
<affiliation wicri:level="3"><inist:fA14 i1="16"><s1>Laboratoire d'Hématologie, CHU</s1>
<s2>Nantes</s2>
<s3>FRA</s3>
<sZ>24 aut.</sZ>
</inist:fA14>
<country>France</country>
<placeName><region type="region">Pays de la Loire</region>
<region type="old region">Pays de la Loire</region>
<settlement type="city">Nantes</settlement>
</placeName>
</affiliation>
</author>
</analytic>
<series><title level="j" type="main">Haematologica : (Roma)</title>
<title level="j" type="abbreviated">Haematologica : (Roma)</title>
<idno type="ISSN">0390-6078</idno>
<imprint><date when="2009">2009</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
<seriesStmt><title level="j" type="main">Haematologica : (Roma)</title>
<title level="j" type="abbreviated">Haematologica : (Roma)</title>
<idno type="ISSN">0390-6078</idno>
</seriesStmt>
</fileDesc>
<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Allele</term>
<term>Concordance</term>
<term>Genetics</term>
<term>Hematology</term>
<term>JAK2 protein tyrosine kinase</term>
<term>Multicenter study</term>
<term>Myeloproliferative syndrome</term>
<term>Polymerase chain reaction</term>
<term>Quantitative analysis</term>
<term>Standardization</term>
</keywords>
<keywords scheme="Pascal" xml:lang="fr"><term>Réaction chaîne polymérase</term>
<term>Concordance</term>
<term>Analyse quantitative</term>
<term>Syndrome myéloprolifératif</term>
<term>Etude multicentrique</term>
<term>Normalisation</term>
<term>Allèle</term>
<term>Génétique</term>
<term>Hématologie</term>
<term>JAK2 protein tyrosine kinase</term>
</keywords>
<keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Analyse quantitative</term>
<term>Normalisation</term>
<term>Génétique</term>
</keywords>
</textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en">Background Many different techniques have been designed for the quantification of JAK2V617F allelic burden, sometimes producing discrepant results. Design and Methods JAK2V617F quantification techniques were compared among 16 centers using 11 assays based on quantitative polymerase chain reaction (with mutation-specific primers or probes, or fluorescent resonance energy transfer/melting curve analysis), allele-specific polymerase chain reaction, conventional sequencing or pyrosequencing. Results A first series of blinded samples (granulocyte DNA, n=29) was analyzed. Seven assays (12 centers) reported values inside the mean±2SD; the mean coefficient of variation was 31%. Sequencing techniques lacked sensitivity, and strong discrepancies were observed with four techniques, which could be attributed to inadequate standards or to different modes of expression of results. Indeed, quantification of JAK2V617F in relation to another control gene produced higher than expected values, suggesting the possibility of more than two JAK2 copies/cell. After calibration of assays with common 1% to 100% JAK2V617F standards (dilutions of UKE-1 cells in normal leukocytes), 14 centers tested ten new samples. JAK2V617F allelic burdens greater or equal than 1% were then reliably quantified by five techniques - one allele specific-polymerase chain reaction and four TaqMan allele-specific quantitative polymerase chain reaction assays, including one previously giving results outside the mean±2SD - with a lower mean coefficient of variation (21%). Of these, only the two TaqMan allele-specific quantitative polymerase chain reaction assays with primer-based specificity could detect 0.2% JAK2V617F. Conclusions Techniques expressing the allelic burden as JAK2V617F/total JAK2 and using a common set of standards produced similar quantification results but with variable sensitivity. Calibration to a reference standard improved reproducibility.</div>
</front>
</TEI>
<affiliations><list><country><li>Allemagne</li>
<li>Australie</li>
<li>France</li>
<li>Italie</li>
<li>Pays-Bas</li>
<li>Royaume-Uni</li>
<li>Suisse</li>
<li>États-Unis</li>
</country>
<region><li>Aquitaine</li>
<li>Auvergne (région administrative)</li>
<li>Auvergne-Rhône-Alpes</li>
<li>Bourgogne</li>
<li>Bourgogne-Franche-Comté</li>
<li>Hambourg</li>
<li>Languedoc-Roussillon</li>
<li>Nouvelle-Aquitaine</li>
<li>Occitanie (région administrative)</li>
<li>Pays de la Loire</li>
<li>Région Bretagne</li>
<li>Texas</li>
<li>Utah</li>
<li>Île-de-France</li>
</region>
<settlement><li>Bordeaux</li>
<li>Brest</li>
<li>Clermont-Ferrand</li>
<li>Dijon</li>
<li>Hambourg</li>
<li>Nantes</li>
<li>Nîmes</li>
<li>Paris</li>
</settlement>
</list>
<tree><country name="France"><region name="Nouvelle-Aquitaine"><name sortKey="Lippert, Eric" sort="Lippert, Eric" uniqKey="Lippert E" first="Eric" last="Lippert">Eric Lippert</name>
</region>
<name sortKey="Carillo, Serge" sort="Carillo, Serge" uniqKey="Carillo S" first="Serge" last="Carillo">Serge Carillo</name>
<name sortKey="Girodon, Francois" sort="Girodon, Francois" uniqKey="Girodon F" first="Francois" last="Girodon">Francois Girodon</name>
<name sortKey="Harrivel, Veronique" sort="Harrivel, Veronique" uniqKey="Harrivel V" first="Véronique" last="Harrivel">Véronique Harrivel</name>
<name sortKey="Hermouet, Sylvie" sort="Hermouet, Sylvie" uniqKey="Hermouet S" first="Sylvie" last="Hermouet">Sylvie Hermouet</name>
<name sortKey="Marzac, Christophe" sort="Marzac, Christophe" uniqKey="Marzac C" first="Christophe" last="Marzac">Christophe Marzac</name>
<name sortKey="Migeon, Marina" sort="Migeon, Marina" uniqKey="Migeon M" first="Marina" last="Migeon">Marina Migeon</name>
<name sortKey="Mounier, Morgane" sort="Mounier, Morgane" uniqKey="Mounier M" first="Morgane" last="Mounier">Morgane Mounier</name>
<name sortKey="Richard, Celine" sort="Richard, Celine" uniqKey="Richard C" first="Céline" last="Richard">Céline Richard</name>
<name sortKey="Ugo, Valerie" sort="Ugo, Valerie" uniqKey="Ugo V" first="Valérie" last="Ugo">Valérie Ugo</name>
</country>
<country name="Australie"><noRegion><name sortKey="Hammond, Emma" sort="Hammond, Emma" uniqKey="Hammond E" first="Emma" last="Hammond">Emma Hammond</name>
</noRegion>
<name sortKey="Herrmann, Richard" sort="Herrmann, Richard" uniqKey="Herrmann R" first="Richard" last="Herrmann">Richard Herrmann</name>
</country>
<country name="États-Unis"><region name="Texas"><name sortKey="Jelinek, Jaroslav" sort="Jelinek, Jaroslav" uniqKey="Jelinek J" first="Jaroslav" last="Jelinek">Jaroslav Jelinek</name>
</region>
<name sortKey="Prchal, Josef T" sort="Prchal, Josef T" uniqKey="Prchal J" first="Josef T." last="Prchal">Josef T. Prchal</name>
<name sortKey="Prchal, Josef T" sort="Prchal, Josef T" uniqKey="Prchal J" first="Josef T." last="Prchal">Josef T. Prchal</name>
<name sortKey="Reading, N Scott" sort="Reading, N Scott" uniqKey="Reading N" first="N. Scott" last="Reading">N. Scott Reading</name>
<name sortKey="Swierczek, Sabina" sort="Swierczek, Sabina" uniqKey="Swierczek S" first="Sabina" last="Swierczek">Sabina Swierczek</name>
</country>
<country name="Allemagne"><region name="Hambourg"><name sortKey="Fehse, Boris" sort="Fehse, Boris" uniqKey="Fehse B" first="Boris" last="Fehse">Boris Fehse</name>
</region>
<name sortKey="Kroger, Nicolaus" sort="Kroger, Nicolaus" uniqKey="Kroger N" first="Nicolaus" last="Kröger">Nicolaus Kröger</name>
</country>
<country name="Royaume-Uni"><noRegion><name sortKey="Hanlon, Katy" sort="Hanlon, Katy" uniqKey="Hanlon K" first="Katy" last="Hanlon">Katy Hanlon</name>
</noRegion>
<name sortKey="Ellard, Sian" sort="Ellard, Sian" uniqKey="Ellard S" first="Sian" last="Ellard">Sian Ellard</name>
</country>
<country name="Pays-Bas"><noRegion><name sortKey="Hermans, Mirjam" sort="Hermans, Mirjam" uniqKey="Hermans M" first="Mirjam" last="Hermans">Mirjam Hermans</name>
</noRegion>
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<country name="Italie"><noRegion><name sortKey="Pietra, Daniela" sort="Pietra, Daniela" uniqKey="Pietra D" first="Daniela" last="Pietra">Daniela Pietra</name>
</noRegion>
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<country name="Suisse"><noRegion><name sortKey="Sobas, Marta" sort="Sobas, Marta" uniqKey="Sobas M" first="Marta" last="Sobas">Marta Sobas</name>
</noRegion>
<name sortKey="Skoda, Radek C" sort="Skoda, Radek C" uniqKey="Skoda R" first="Radek C." last="Skoda">Radek C. Skoda</name>
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Concordance of assays designed for the quantification of JAK2V617F : a multicenter study

Concordance of assays designed for the quantification of JAK2V617F : a multicenter study

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