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

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Concordance of assays designed for the quantification of JAK2V617F: a multicenter study. / Lippert, Eric; Girodon, François; Hammond, Emma; Jelinek, Jaroslav; Reading, N Scott; Fehse, Boris; Hanlon, Katy; Hermans, Mirjam; Richard, Céline; Swierczek, Sabina; Ugo, Valérie; Carillo, Serge; Harrivel, Véronique; Marzac, Christophe; Pietra, Daniela; Sobas, Marta; Mounier, Morgane; Migeon, Marina; Ellard, Sian; Kröger, Nicolaus; Herrmann, Richard; Prchal, Josef T; Skoda, Radek C; Hermouet, Sylvie.

In: HAEMATOLOGICA, Vol. 94, No. 1, 1, 2009, p. 38-45.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Lippert, E, Girodon, F, Hammond, E, Jelinek, J, Reading, NS, Fehse, B, Hanlon, K, Hermans, M, Richard, C, Swierczek, S, Ugo, V, Carillo, S, Harrivel, V, Marzac, C, Pietra, D, Sobas, M, Mounier, M, Migeon, M, Ellard, S, Kröger, N, Herrmann, R, Prchal, JT, Skoda, RC & Hermouet, S 2009, 'Concordance of assays designed for the quantification of JAK2V617F: a multicenter study.', HAEMATOLOGICA, vol. 94, no. 1, 1, pp. 38-45. https://doi.org/10.3324/haematol.13486

APA

Lippert, E., Girodon, F., Hammond, E., Jelinek, J., Reading, N. S., Fehse, B., Hanlon, K., Hermans, M., Richard, C., Swierczek, S., Ugo, V., Carillo, S., Harrivel, V., Marzac, C., Pietra, D., Sobas, M., Mounier, M., Migeon, M., Ellard, S., ... Hermouet, S. (2009). Concordance of assays designed for the quantification of JAK2V617F: a multicenter study. HAEMATOLOGICA, 94(1), 38-45. [1]. https://doi.org/10.3324/haematol.13486

Vancouver

Lippert E, Girodon F, Hammond E, Jelinek J, Reading NS, Fehse B et al. Concordance of assays designed for the quantification of JAK2V617F: a multicenter study. HAEMATOLOGICA. 2009;94(1):38-45. 1. https://doi.org/10.3324/haematol.13486

Bibtex

@article{8807d64f45b34cbc95b49cd1e7a3f794,
title = "Concordance of assays designed for the quantification of JAK2V617F: a multicenter study.",
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.",
author = "Eric Lippert and Fran{\c c}ois Girodon and Emma Hammond and Jaroslav Jelinek and Reading, {N Scott} and Boris Fehse and Katy Hanlon and Mirjam Hermans and C{\'e}line Richard and Sabina Swierczek and Val{\'e}rie Ugo and Serge Carillo and V{\'e}ronique Harrivel and Christophe Marzac and Daniela Pietra and Marta Sobas and Morgane Mounier and Marina Migeon and Sian Ellard and Nicolaus Kr{\"o}ger and Richard Herrmann and Prchal, {Josef T} and Skoda, {Radek C} and Sylvie Hermouet",
year = "2009",
doi = "10.3324/haematol.13486",
language = "Deutsch",
volume = "94",
pages = "38--45",
journal = "HAEMATOLOGICA",
issn = "0390-6078",
publisher = "Ferrata Storti Foundation",
number = "1",

}

RIS

TY - JOUR

T1 - Concordance of assays designed for the quantification of JAK2V617F: a multicenter study.

AU - Lippert, Eric

AU - Girodon, François

AU - Hammond, Emma

AU - Jelinek, Jaroslav

AU - Reading, N Scott

AU - Fehse, Boris

AU - Hanlon, Katy

AU - Hermans, Mirjam

AU - Richard, Céline

AU - Swierczek, Sabina

AU - Ugo, Valérie

AU - Carillo, Serge

AU - Harrivel, Véronique

AU - Marzac, Christophe

AU - Pietra, Daniela

AU - Sobas, Marta

AU - Mounier, Morgane

AU - Migeon, Marina

AU - Ellard, Sian

AU - Kröger, Nicolaus

AU - Herrmann, Richard

AU - Prchal, Josef T

AU - Skoda, Radek C

AU - Hermouet, Sylvie

PY - 2009

Y1 - 2009

N2 - 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.

AB - 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.

U2 - 10.3324/haematol.13486

DO - 10.3324/haematol.13486

M3 - SCORING: Zeitschriftenaufsatz

VL - 94

SP - 38

EP - 45

JO - HAEMATOLOGICA

JF - HAEMATOLOGICA

SN - 0390-6078

IS - 1

M1 - 1

ER -