目的:对国产血浆游离DNA(cell-free DNA,cfDNA)提取试剂盒进行评价。方法:用志愿者血浆构成血浆池A,再分别加入固定量片段长度为102、268、402 bp和所有片段混合物的DNA,形成血浆池B1-B4。用Qiagen和国产柱式法cfDNA提取试剂盒分别提取血浆池A和血浆池B1-B4中的cfDNA。用Taq Man qPCR分析两种试剂盒提取血浆池A中cfDNA的提取效率,同时分析cfDNA浓度与血浆用量之间的关系,用琼脂糖凝胶电泳分析提取的cfDNA的片段完整性。核酸测定仪分析两种试剂盒对血浆池B1-B4中不同DNA片段的回收率。结果:Qiagen和国产试剂盒提取的血浆池A中cfDNA的Ct值分别为27.94±0.423和27.41±0.356,国产试剂盒提取效率明显高于Qiagen试剂盒(t=4.29,P<0.01);琼脂糖凝胶电泳结果表明,两种方法提取的血浆池A中cfDNA在102、268和402 bp处均出现明显条带,而1 204 bp片段未出现。Qiagen和国产试剂盒提取的cfDNA浓度与血浆池A的血浆用量之间均存在明显的线性相关,r~2分别为0.979(P<0.01)和0.96...
【英文摘要】
Objective: To evaluate the domestic kit of cell-free DNA( cfDNA) extraction from plasma. Methods: Plasma pool A was composed of volunteers plasma,and different length of DNAs were added into the plasma pool A to form the plasma pool B1-B4,the addition of DNA fragments length were 102,268 and 402 bp and their mixture,respectively. The cfDNA in plasma pool A and pool B1-B4 were extracted by Qiagen and domestic column free DNA extraction kit. The extraction efficiencies of cfDNA in plasma pool A of two Extract...
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