Abstract：
Genotoxicity is an important information that should be included in human biomonitoring programmes. However, the usually applied cytogenetic assays are laborious and time-consuming, reason why it is critical to develop rapid and economic new methods. The aim of this study was to evaluate if the molecular profile of frozen whole blood, acquired by Fourier Transform Infrared (FTIR) spectroscopy, allows to assess genotoxicity in occupational exposure to antineoplastic drugs, as obtained by the cytokinesis-block micronucleus assay. For that purpose, 92 samples of peripheral blood were studied: 46 samples from hospital professionals occupationally exposed to antineoplastic drugs and 46 samples from workers in academia without exposure (controls). It was first evaluated the metabolome from frozen whole blood by methanol precipitation of macromolecules as haemoglobin, followed by centrifugation. The metabolome molecular profile resulted in 3 ratios of spectral bands, significantly different between the exposed and non-exposed group (p < 0.01) and a spectral principal component-linear discriminant analysis (PCA-LDA) model enabling to predict genotoxicity from exposure with 73 % accuracy. After optimization of the dilution degree and solution used, it was possible to obtain a higher number of significant ratios of spectral bands, i.e., 10 ratios significantly different (p < 0.001), highlighting the high sensitivity and specificity of the method. Indeed, the PCA-LDA model, based on the molecular profile of whole blood, enabled to predict genotoxicity from the exposure with an accuracy, sensitivity, and specificity of 92 %, 93 % and 91 %, respectively. All these parameters were achieved based on 1 μL of frozen whole blood, in a high-throughput mode, i.e., based on the simultaneous analysis of 92 samples, in a simple and economic mode. In summary, it can be conclude that this method presents a very promising potential for high-dimension screening of exposure to genotoxic substances.
摘要：
遗传毒性是人类生物监测计划中应包括的重要信息。然而,通常应用的细胞遗传学测定是费力和耗时的，发展快速经济的新方法至关重要的原因。这项研究的目的是评估冷冻全血的分子谱，傅里叶变换红外(FTIR)光谱,允许评估职业性接触抗肿瘤药物的遗传毒性，通过胞质分裂阻滞微核试验获得。为此,研究了92个外周血样本：来自职业暴露于抗肿瘤药物的医院专业人员的46个样本和来自学术界未暴露的46个样本（对照）。首先通过将大分子甲醇沉淀为血红蛋白来评估冷冻全血的代谢组，然后离心。代谢组分子谱导致3个光谱带比率，暴露组和非暴露组之间存在显着差异（p<0.01），并且光谱主成分线性判别分析（PCA-LDA）模型能够以73％的准确度预测暴露的遗传毒性。在优化稀释度和使用的溶液后，有可能获得更多的光谱带显著比率，即，10个比率显著不同(p<0.001)，突出了该方法的高灵敏度和特异性。的确,PCA-LDA模型，根据全血的分子特征,能够准确预测暴露的遗传毒性，灵敏度,和92%的特异性，93%和91%，分别。所有这些参数都是基于1μL冷冻全血获得的，在高吞吐量模式下，即，基于对92个样本的同时分析，以简单经济的方式。总之,可以得出结论，这种方法对于高维筛查遗传毒性物质的暴露具有非常有希望的潜力。
