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Clin Chem:合成循环游离DNA作为癌症液体活检的体细胞突变检测的质控材料

2017-08-29 MedSci MedSci原创

研究人员开发了一套合成cfDNA质控材料(SCQCM),在微球菌核酸酶消化系统的基础上,其携带体细胞突变作为模拟cfDNA并且能够与基因组DNA匹配作为遗传背景,以模拟真实临床中的配对肿瘤标准样品试验。使用来自CRISPR / Cas9编辑的具有EML4-ALK重排的细胞的单核苷酸变体或indel和基因组DNA含有1500-2000个碱基的定点诱变被片段化,定量并加入到衍生自HEK293T细胞的微

近日,国际杂志 《Clinical Chemistry》上在线发表一项关于合成循环游离DNA作为癌症液体活检的体细胞突变检测的质控材料的研究。

由于cfDNA浓度低、检测方法多变、工作流程复杂,在血浆中检测肿瘤衍生细胞游离DNA(cfDNA)的体细胞基因组改变仍然具有很大困难。此外,目前还没有合适的质控材料。

研究人员开发了一套合成cfDNA质控材料(SCQCM),在微球菌核酸酶消化系统的基础上,其携带体细胞突变作为模拟cfDNA并且能够与基因组DNA匹配作为遗传背景,以模拟真实临床中的配对肿瘤标准样品试验。使用来自CRISPR / Cas9编辑的具有EML4-ALK重排的细胞的单核苷酸变体或indel和基因组DNA含有1500-2000个碱基的定点诱变被片段化,定量并加入到衍生自HEK293T细胞的微球菌核酸酶消化DNA中。为了证明其适用性,将SCQCM与患者衍生的血浆样品进行比较,并在包含11个实验室的合作研究中进行了验证。

下一代测序的SCQCM分析结果与患者血浆样品的结果一致,包括cfDNA的大小分布和测序数据的质量控制指标。超过95%的实验室正确检测到EGFR T790ML858RKRAS G12D和外显子19以及EML4-ALK变体2中的缺失的SCQCM

研究认为,SCQCMs可成功应用于更为广泛的环境,此外,SCQCM可以用作循环肿瘤DNA体细胞突变检测的测试性能评估中的最佳质控材料。

原始出处:

Rui Zhang, Rongxue Peng, Ziyang Li,et al. Synthetic Circulating Cell-free DNA as Quality Control Materials for Somatic Mutation Detection in Liquid Biopsy for Cancer

本文系梅斯医学(MedSci)原创编译整理,转载需授权!


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    2017-08-31 smlt2008
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    2017-08-31 LSJ116

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