A new biodosimetric method: branched DNA-based quantitative detection of B1 DNA in mouse plasma.
A new biodosimetric method: branched DNA-based quantitative detection of B1 DNA in mouse plasma.
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DOI：
10.1259/bjr/49886569
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发表时间：
2010-08
期刊：
影响因子：
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通讯作者：
L. Zhang;M. Zhang;S. Yang;Y. Cao;S. Bingrong Zhang;L. Yin;Y. Tian;Y. Ma;A. Zhang;P. Okunieff;L. Zhang
中科院分区：
文献类型：
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作者：
L. Zhang;M. Zhang;S. Yang;Y. Cao;S. Bingrong Zhang;L. Yin;Y. Tian;Y. Ma;A. Zhang;P. Okunieff;L. Zhang
一个简单而准确的方法来测量辐射的生物效应越来越重要，特别是在大规模伤亡的情况下。因此，我们开发了一种新的生物剂量测定技术，针对循环B1 DNA在小鼠血浆中的分支DNA信号放大的血浆DNA的快速定量。该技术靶向小鼠基因组中B1反转录转座子的重复元件，然后使用Panomics Quantigene 2.0试剂进行信号扩增。对精密度、准确度和线性进行了评价。在0-10戈伊全身照射（TBI）后0-24 h从小鼠收集血浆样品。平均批间和批内变异系数分别为8.7%和12.3%。血浆加标DNA的平均回收率为89.5%。结果表明，BALB/c小鼠和NIH Swiss小鼠在6戈伊TBI后，血浆B1 DNA水平在3 h显著升高，9 h达高峰，24 h逐渐恢复至基线水平。在TBI后9 h观察到血浆DNA的剂量依赖性变化;剂量-反应关系是单调的，BALB/c小鼠在3 - 6戈伊（r = 0.993）和NIH Swiss小鼠在3 - 7戈伊（r = 0.98）呈现线性。这种基于分支DNA的检测方法在定量检测血浆B1 DNA方面可靠、准确、灵敏。在3 ~ 6戈伊剂量范围内，BALB/c和NIH Swiss小鼠血浆B1 DNA呈辐射剂量相关性增加，表明血浆B1 DNA有可能作为辐射生物学效应的生物标志物。
A simple and accurate method for measuring the biological effects of radiation is of increasing importance, especially in mass casualty scenarios. We have therefore developed a new biodosimetric technique targeting circulating B1 DNA in mouse plasma by branched DNA signal amplification for rapid quantification of plasma DNA. This technology targets repetitive elements of the B1 retrotransposon in the mouse genome, followed by signal amplification using Panomics Quantigene 2.0 reagents. Evaluation was conducted concerning precision, accuracy and linearity. Plasma samples were collected from mice 0-24 h after 0-10 Gy total body irradiation (TBI). The average inter- and intra-assay coefficients of variance were 8.7% and 12.3%, respectively. The average recovery rate of spiked DNA into plasma was 89.5%. This assay revealed that when BALB/c and NIH Swiss mice were exposed to 6 Gy TBI, plasma B1 DNA levels increased significantly at 3 h post-TBI, peaked at 9 h and gradually returned toward baseline levels in 24 h. A dose-dependent change in plasma DNA was observed at 9 h post-TBI; the dose-response relation was monotonic, exhibiting linearity for BALB/c mice from 3 to 6 Gy (r = 0.993) and NIH Swiss mice from 3 to 7 Gy (r = 0.98). This branched DNA-based assay is reliable, accurate and sensitive in detecting plasma B1 DNA quantitatively. A radiation dose-correlated increase in plasma B1 DNA was demonstrated in BALB/c and NIH Swiss mice in the dose range from 3 to 6 Gy, suggesting that plasma B1 DNA has potential as a biomarker for radiation biological effect.