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

作者：Rothkamm Kai^*; Barnard Stephen; Ain**ury Elizabeth A; Al hafidh Jenna; Barquinero Joan Francesc; Lindholm Carita; Moquet Jayne; Perala Marjo; Roch Lefevre Sandrine; Scherthan Harry; Thierens Hubert; Vral Anne; Vandersickel Veerle

来源：Mutation Research: Genetic Toxicology and Environmental Mutagenesis , 2013, 756(1-2): 170-173.

DOI：10.1016/j.mrgentox.2013.04.012

• Biological dosimetry
• Gamma-H2AX
• Radiation-induced foci
• Ionizing radiation
• DNA double-strand break
• Biomarker

The identification of severely exposed individuals and reassurance of the %26apos;worried well%26apos; are of prime importance for initial triage following a large scale radiation accident. We aim to develop the gamma-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 gamma-irradiated, 4 or 24 h incubated and overnight-shipped lymphocyte samples from four donors to generate gamma-H2AX reference data, using manual and/or automated foci scoring strategies. In addition to acute, homogeneous exposures to 0, 1, 2 and 4 Gy, acute simulated partial body (4 Gy to 50% of cells) and protracted exposures (4 Gy over 24 h) were analysed. Data from all laboratories could be satisfactorily fitted with linear dose response functions. Average yields observed at 4 h post exposure were 2-4 times higher than at 24 h 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 gamma-H2AX assay. We conclude that the gamma-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.

• 出版日期2013-8-30