Manual versus automated γ-H2AX foci analysis across five European laboratories: can this assay be used for rapid biodosimetry in a large scale radiation accident?

Kai Rothkamm; Stephen Barnard; Elizabeth A Ainsbury; Jenna Al-Hafidh; Joan-Francesc Barquinero; Carita Lindholm; Jayne Moquet; Marjo Perälä; Sandrine Roch-Lefèvre; Harry Scherthan; Hubert Thierens; Anne Vral; Veerle Vandersickel

doi:10.1016/j.mrgentox.2013.04.012

Manual versus automated γ-H2AX foci analysis across five European laboratories

Standard

Manual versus automated γ-H2AX foci analysis across five European laboratories : can this assay be used for rapid biodosimetry in a large scale radiation accident? / Rothkamm, Kai; Barnard, Stephen; Ainsbury, Elizabeth A; Al-Hafidh, Jenna; Barquinero, Joan-Francesc; Lindholm, Carita; Moquet, Jayne; Perälä, Marjo; Roch-Lefèvre, Sandrine; Scherthan, Harry; Thierens, Hubert; Vral, Anne; Vandersickel, Veerle.

In: MUTAT RES-FUND MOL M, Vol. 756, No. 1-2, 30.08.2013, p. 170-3.

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

Harvard

Rothkamm, K, Barnard, S, Ainsbury, EA, Al-Hafidh, J, Barquinero, J-F, Lindholm, C, Moquet, J, Perälä, M, Roch-Lefèvre, S, Scherthan, H, Thierens, H, Vral, A & Vandersickel, V 2013, 'Manual versus automated γ-H2AX foci analysis across five European laboratories: can this assay be used for rapid biodosimetry in a large scale radiation accident?', MUTAT RES-FUND MOL M, vol. 756, no. 1-2, pp. 170-3. https://doi.org/10.1016/j.mrgentox.2013.04.012

APA

Rothkamm, K., Barnard, S., Ainsbury, E. A., Al-Hafidh, J., Barquinero, J-F., Lindholm, C., Moquet, J., Perälä, M., Roch-Lefèvre, S., Scherthan, H., Thierens, H., Vral, A., & Vandersickel, V. (2013). Manual versus automated γ-H2AX foci analysis across five European laboratories: can this assay be used for rapid biodosimetry in a large scale radiation accident? MUTAT RES-FUND MOL M, 756(1-2), 170-3. https://doi.org/10.1016/j.mrgentox.2013.04.012

Vancouver

Rothkamm K, Barnard S, Ainsbury EA, Al-Hafidh J, Barquinero J-F, Lindholm C et al. Manual versus automated γ-H2AX foci analysis across five European laboratories: can this assay be used for rapid biodosimetry in a large scale radiation accident? MUTAT RES-FUND MOL M. 2013 Aug 30;756(1-2):170-3. https://doi.org/10.1016/j.mrgentox.2013.04.012

Bibtex

@article{7870e3c79d654e659f87ab1965c9ae22,

title = "Manual versus automated γ-H2AX foci analysis across five European laboratories: can this assay be used for rapid biodosimetry in a large scale radiation accident?",

abstract = "The identification of severely exposed individuals and reassurance of the 'worried well' are of prime importance for initial triage following a large scale radiation accident. We aim to develop the γ-H2AX foci assay into a rapid biomarker tool for use in accidents. Here, five laboratories established a standard operating procedure and analysed 100 ex vivo γ-irradiated, 4 or 24h incubated and overnight-shipped lymphocyte samples from four donors to generate γ-H2AX reference data, using manual and/or automated foci scoring strategies. In addition to acute, homogeneous exposures to 0, 1, 2 and 4Gy, acute simulated partial body (4Gy to 50% of cells) and protracted exposures (4Gy over 24h) were analysed. Data from all laboratories could be satisfactorily fitted with linear dose response functions. Average yields observed at 4h post exposure were 2-4 times higher than at 24h and varied considerably between laboratories. Automated scoring caused larger uncertainties than manual scoring and was unable to identify partial exposures, which were detectable in manually scored samples due to their overdispersed foci distributions. Protracted exposures were detectable but doses could not be accurately estimated with the γ-H2AX assay. We conclude that the γ-H2AX assay may be useful for rapid triage following a recent acute radiation exposure. The potentially higher speed and convenience of automated relative to manual foci scoring needs to be balanced against its compromised accuracy and inability to detect partial body exposures. Regular re-calibration or inclusion of reference samples may be necessary to ensure consistent results between laboratories or over long time periods. ",

keywords = "Automation, Dose-Response Relationship, Radiation, Europe, Gamma Rays/adverse effects, Histones/analysis, Humans, Laboratories/standards, Lymphocytes/metabolism, Microscopy, Fluorescence, Radiation Monitoring/methods, Radioactive Hazard Release/prevention & control, Time Factors",

author = "Kai Rothkamm and Stephen Barnard and Ainsbury, {Elizabeth A} and Jenna Al-Hafidh and Joan-Francesc Barquinero and Carita Lindholm and Jayne Moquet and Marjo Per{\"a}l{\"a} and Sandrine Roch-Lef{\`e}vre and Harry Scherthan and Hubert Thierens and Anne Vral and Veerle Vandersickel",

year = "2013",

month = aug,

day = "30",

doi = "10.1016/j.mrgentox.2013.04.012",

language = "English",

volume = "756",

pages = "170--3",

number = "1-2",

}

RIS

TY - JOUR

T1 - Manual versus automated γ-H2AX foci analysis across five European laboratories

T2 - can this assay be used for rapid biodosimetry in a large scale radiation accident?

AU - Rothkamm, Kai

AU - Barnard, Stephen

AU - Ainsbury, Elizabeth A

AU - Al-Hafidh, Jenna

AU - Barquinero, Joan-Francesc

AU - Lindholm, Carita

AU - Moquet, Jayne

AU - Perälä, Marjo

AU - Roch-Lefèvre, Sandrine

AU - Scherthan, Harry

AU - Thierens, Hubert

AU - Vral, Anne

AU - Vandersickel, Veerle

PY - 2013/8/30

Y1 - 2013/8/30

N2 - The identification of severely exposed individuals and reassurance of the 'worried well' are of prime importance for initial triage following a large scale radiation accident. We aim to develop the γ-H2AX foci assay into a rapid biomarker tool for use in accidents. Here, five laboratories established a standard operating procedure and analysed 100 ex vivo γ-irradiated, 4 or 24h incubated and overnight-shipped lymphocyte samples from four donors to generate γ-H2AX reference data, using manual and/or automated foci scoring strategies. In addition to acute, homogeneous exposures to 0, 1, 2 and 4Gy, acute simulated partial body (4Gy to 50% of cells) and protracted exposures (4Gy over 24h) were analysed. Data from all laboratories could be satisfactorily fitted with linear dose response functions. Average yields observed at 4h post exposure were 2-4 times higher than at 24h and varied considerably between laboratories. Automated scoring caused larger uncertainties than manual scoring and was unable to identify partial exposures, which were detectable in manually scored samples due to their overdispersed foci distributions. Protracted exposures were detectable but doses could not be accurately estimated with the γ-H2AX assay. We conclude that the γ-H2AX assay may be useful for rapid triage following a recent acute radiation exposure. The potentially higher speed and convenience of automated relative to manual foci scoring needs to be balanced against its compromised accuracy and inability to detect partial body exposures. Regular re-calibration or inclusion of reference samples may be necessary to ensure consistent results between laboratories or over long time periods.

AB - The identification of severely exposed individuals and reassurance of the 'worried well' are of prime importance for initial triage following a large scale radiation accident. We aim to develop the γ-H2AX foci assay into a rapid biomarker tool for use in accidents. Here, five laboratories established a standard operating procedure and analysed 100 ex vivo γ-irradiated, 4 or 24h incubated and overnight-shipped lymphocyte samples from four donors to generate γ-H2AX reference data, using manual and/or automated foci scoring strategies. In addition to acute, homogeneous exposures to 0, 1, 2 and 4Gy, acute simulated partial body (4Gy to 50% of cells) and protracted exposures (4Gy over 24h) were analysed. Data from all laboratories could be satisfactorily fitted with linear dose response functions. Average yields observed at 4h post exposure were 2-4 times higher than at 24h and varied considerably between laboratories. Automated scoring caused larger uncertainties than manual scoring and was unable to identify partial exposures, which were detectable in manually scored samples due to their overdispersed foci distributions. Protracted exposures were detectable but doses could not be accurately estimated with the γ-H2AX assay. We conclude that the γ-H2AX assay may be useful for rapid triage following a recent acute radiation exposure. The potentially higher speed and convenience of automated relative to manual foci scoring needs to be balanced against its compromised accuracy and inability to detect partial body exposures. Regular re-calibration or inclusion of reference samples may be necessary to ensure consistent results between laboratories or over long time periods.

KW - Automation

KW - Dose-Response Relationship, Radiation

KW - Europe

KW - Gamma Rays/adverse effects

KW - Histones/analysis

KW - Humans

KW - Laboratories/standards

KW - Lymphocytes/metabolism

KW - Microscopy, Fluorescence

KW - Radiation Monitoring/methods

KW - Radioactive Hazard Release/prevention & control

KW - Time Factors

U2 - 10.1016/j.mrgentox.2013.04.012

DO - 10.1016/j.mrgentox.2013.04.012

M3 - SCORING: Journal article

C2 - 23648320

VL - 756

SP - 170

EP - 173

IS - 1-2

ER -