如何提高 NGS 方案对于低频突变信号的检出率？
① 增加样本投入量：样本起始量越多，支持低频突变检出的模板拷贝数越多；
② 增加跟踪突变数量：突变位点数量越高，检出概率越高。
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Van Der Pol Y, Mouliere F. Toward the early detection of cancer by decoding the epigenetic and environmental fingerprints of cell-free DNA[J]. Cancer cell, 2019, 36(4): 350-368.
不同实验条件下对应的突变检测下限
|
文库转化率 |
样本起始量 |
|||||||||
|
1 ng |
10 ng |
20 ng |
30 ng |
50 ng |
100 ng |
200 ng |
300 ng |
500 ng |
1000 ng |
|
|
20% |
5.000% |
0.500% |
0.250% |
0.168% |
0.100% |
0.050% |
0.025% |
0.017% |
0.010% |
0.005% |
|
30% |
3.400% |
0.340% |
0.170% |
0.113% |
0.068% |
0.034% |
0.017% |
0.011% |
0.007% |
0.003% |
|
50% |
2.000% |
0.200% |
0.100% |
0.067% |
0.040% |
0.020% |
0.010% |
0.007% |
0.004% |
0.002% |
|
100% |
1.000% |
0.100% |
0.050% |
0.033% |
0.020% |
0.010% |
0.005% |
0.003% |
0.002% |
0.001% |
当文库转化率为 50% 时，投入 400 ng 的样本可检出 0.005% 的突变；
② 增加突变位点数量