Romm H, Ainsbury E, Bajinskis A, Barnard S, Barquinero J F, Barrios L, Beinke C, Puig-Casanovas R, Deperas-Kaminska M, Gregoire E, Oestreicher U, Lindholm C, Moquet J, Rothkamm K, Sommer S, Thierens H, Vral A, Vandersickel V, Wojcik A
Bundesamt fuer Strahlenschutz, Ingolstaedter Landstr. 1, 85764, Neuherberg, Germany,
Radiat Environ Biophys. 2014 May;53(2):241-54. doi: 10.1007/s00411-014-0519-8. Epub 2014 Feb 21.
In the case of a large scale radiation accident high throughput methods of biological dosimetry for population triage are needed to identify individuals requiring clinical treatment. The dicentric assay performed in web-based scoring mode may be a very suitable technique. Within the MULTIBIODOSE EU FP7 project a network is being established of 8 laboratories with expertise in dose estimations based on the dicentric assay. Here, the manual dicentric assay was tested in a web-based scoring mode. More than 23,000 high resolution images of metaphase spreads (only first mitosis) were captured by four laboratories and established as image galleries on the internet (cloud). The galleries included images of a complete dose effect curve (0-5.0 Gy) and three types of irradiation scenarios simulating acute whole body, partial body and protracted exposure. The blood samples had been irradiated in vitro with gamma rays at the University of Ghent, Belgium. Two laboratories provided image galleries from Fluorescence plus Giemsa stained slides (3 h colcemid) and the image galleries from the other two laboratories contained images from Giemsa stained preparations (24 h colcemid). Each of the 8 participating laboratories analysed 3 dose points of the dose effect curve (scoring 100 cells for each point) and 3 unknown dose points (50 cells) for each of the 3 simulated irradiation scenarios. At first all analyses were performed in a QuickScan Mode without scoring individual chromosomes, followed by conventional scoring (only complete cells, 46 centromeres). The calibration curves obtained using these two scoring methods were very similar, with no significant difference in the linear-quadratic curve coefficients. Analysis of variance showed a significant effect of dose on the yield of dicentrics, but no significant effect of the laboratories, different methods of slide preparation or different incubation times used for colcemid. The results obtained to date within the MULTIBIODOSE project by a network of 8 collaborating laboratories throughout Europe are very promising. The dicentric assay in the web based scoring mode as a high throughput scoring strategy is a useful application for biodosimetry in the case of a large scale radiation accident.
在大规模辐射事故的情况下，需要用于人群分类的生物剂量测定高通量方法来识别需要临床治疗的个体。以基于网络的评分模式进行的双着丝粒分析可能是一种非常合适的技术。在MULTIBIODOSE欧盟第七框架项目中，正在建立一个由8个实验室组成的网络，这些实验室在基于双着丝粒分析的剂量估计方面具有专业知识。在此，对基于网络评分模式下的手工双着丝粒分析进行了测试。四个实验室捕获了超过23000张中期分裂相（仅第一次有丝分裂）的高分辨率图像，并在互联网（云端）上建立了图像库。这些图库包括完整剂量效应曲线（0 - 5.0 Gy）的图像以及模拟急性全身、局部身体和长期照射的三种照射场景类型。血液样本在比利时根特大学进行了体外伽马射线照射。两个实验室提供了荧光加吉姆萨染色玻片（秋水仙酰胺处理3小时）的图像库，另外两个实验室的图像库包含吉姆萨染色制剂（秋水仙酰胺处理24小时）的图像。8个参与实验室中的每个实验室都分析了剂量效应曲线的3个剂量点（每个点对100个细胞进行评分）以及3种模拟照射场景中每种场景的3个未知剂量点（50个细胞）。首先，所有分析均在快速扫描模式下进行，不对单个染色体进行评分，随后进行常规评分（仅对完整细胞，46个着丝粒）。使用这两种评分方法获得的校准曲线非常相似，线性 - 二次曲线系数无显著差异。方差分析表明剂量对双着丝粒产率有显著影响，但实验室、不同的玻片制备方法或秋水仙酰胺使用的不同孵育时间无显著影响。到目前为止，整个欧洲8个合作实验室组成的网络在MULTIBIODOSE项目中获得的结果非常有前景。基于网络评分模式的双着丝粒分析作为一种高通量评分策略，在大规模辐射事故情况下对于生物剂量测定是一种有用的应用。
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