Johnston Matthew L, Young Erik F, Shepard Kenneth L
Bialanx, Inc., 511 Avenue of the Americas, Suite 267, New York, NY, USA,
Radiat Environ Biophys. 2015 Aug;54(3):365-72. doi: 10.1007/s00411-015-0595-4. Epub 2015 May 3.
The current state of the art in high-throughput minimally invasive radiation biodosimetry involves the collection of samples in the field and analysis at a centralized facility. We have developed a simple biological immunoassay for radiation exposure that could extend this analysis out of the laboratory into the field. Such a forward placed assay would facilitate triage of a potentially exposed population. The phosphorylation and localization of the histone H2AX at double-stranded DNA breaks has already been proven to be an adequate surrogate assay for reporting DNA damage proportional to radiation dose. Here, we develop an assay for phosphorylated H2AX directed against minimally processed sample lysates. We conduct preliminary verification of H2AX phosphorylation using irradiated mouse embryo fibroblast cultures. Additional dosimetry is performed using human blood samples irradiated ex vivo. The assay reports H2AX phosphorylation in human blood samples in response to ionizing radiation over a range of 0-5 Gy in a linear fashion, without requiring filtering, enrichment, or purification of the blood sample.
高通量微创辐射生物剂量测定的当前技术水平涉及在现场采集样本并在集中设施进行分析。我们开发了一种用于辐射暴露的简单生物免疫测定法，该方法可将这种分析从实验室扩展到现场。这种前置测定法将有助于对潜在暴露人群进行分类。组蛋白H2AX在双链DNA断裂处的磷酸化和定位已被证明是一种合适的替代测定法，用于报告与辐射剂量成比例的DNA损伤。在此，我们开发了一种针对最少处理的样品裂解物的磷酸化H2AX测定法。我们使用受辐照的小鼠胚胎成纤维细胞培养物对H2AX磷酸化进行初步验证。使用离体辐照的人类血液样本进行额外的剂量测定。该测定法以线性方式报告人类血液样本中H2AX的磷酸化，以响应0 - 5 Gy范围内的电离辐射，无需对血液样本进行过滤、富集或纯化。
