中文摘要：

为了实现对核酸的高灵敏度检测,构建了一种新型的液滴式数字聚合酶链式反应（dd PCR）芯片.该芯片由产生液滴的聚二甲基硅氧烷（PDMS）模块和储存液滴的玻璃腔室构成.实验结果表明,该芯片可以在25 min内产生2×106个直径为20μm的微液滴（体积4.187 p L）.利用该芯片定量检测了表皮生长因子受体（EGFR）基因第19号外显子,在DNA浓度为106~101copies/μL范围内呈现良好的线性关系（R2=0.9998）;在浓度为106copies/μL的19号外显子野生型DNA中检测105~100copies/μL的突变型DNA,其检测敏感度可达到0.0001%.该方法在同一芯片上实现了液滴产生、核酸扩增和荧光信号读取的功能,在核酸绝对定量及痕量突变基因的检测中具有潜在应用前景.

英文摘要：

Sensitive and rapid quantification of nucleic acid is significant for clinical diagnosis and medication guide of tumor. In this study, we developed a novel droplet digital PCR（ddPCR） device for sensitive nucleic acid detection. This chip contains a PDMS chip and glass-made reservoir, which can overcome the problems of droplet evaporation and instability during PCR. The double droplet generation channels in this chip can produce 2×106 droplets（4. 187 pL each） in 25 min. For detecting EGFR exon 19 wild-type DNA using this ddPCR chip, it demonstrated a good linear correlation with the DNA concentration from 10 to 106 copies/μL （R2= 0. 9998）. In addition, we show sensitive detection of EGFR exon 19 mutate DNA in a 106-fold excess of wild-type bac PCR process nucleic acids kground. In conclusion, this ddPCR chip integrates the functions of droplet generation, on-chip and fluorescence signals readout, which will provide a potential application in the absolute quantification and rare mutate gene detection.