Hu Wei, Chen Rong-hua, Zhang Chen, An Zhi-yuan, Wang Bing, Ping Yuan
Fa Yi Xue Za Zhi. 2014 Oct;30(5):342-5.
To test droplet digital PCR for species identification and absolute quantification of biological sample.
Specific primers and probes for human mtDNA encoding gene ND4 and 16S rRNA were designed, and the species-specificity was assessed on DNA samples derived from human and common animals. To determine the sensitivity and stability of droplet digital PCR for species identification and absolute quantification, gradient dilution series of recombinant plasmid and 16 human DNA samples were analyzed.
Human recombinant plasmid FAM (ND4) could be used in detecting the samples of human. And the results of detecting were consistent with all levels of diluted concentrations. Droplet digital PCR was able to detect low and single copy of target DNA.
Droplet digital PCR, with high sensitivity and specificity, is fully amenable for species identification and absolute quantification of biological samples, also it can be applied on routine forensic examination.
测试液滴数字PCR用于生物样本的物种鉴定和绝对定量。
设计针对人类线粒体DNA编码基因ND4和16S rRNA的特异性引物和探针，并对来自人类和常见动物的DNA样本进行物种特异性评估。为了确定液滴数字PCR用于物种鉴定和绝对定量的灵敏度和稳定性，分析了重组质粒的梯度稀释系列和16个人类DNA样本。
人类重组质粒FAM（ND4）可用于检测人类样本。检测结果与所有稀释浓度水平一致。液滴数字PCR能够检测低拷贝和单拷贝的目标DNA。
液滴数字PCR具有高灵敏度和特异性，完全适用于生物样本的物种鉴定和绝对定量，也可应用于常规法医检验。
