分析化学2025，Vol.53Issue(7)：1138-1145,8.DOI:10.19756/j.issn.0253-3820.251066
双体积液滴对数字液滴PCR定量检测能力提升的研究
Study on Improvement of Quantitative Capacity of Digital Droplet PCR by Double-Volume Droplets
摘要
Abstract
Digital polymerase chain reaction(dPCR)enables absolute quantitative detection of nucleic acid samples.Since the quantitative upper and lower limits of detectable samples mainly depend on the volume and number of single droplets,the abundance of the sample to be tested,the volume of single droplets,and the number of droplets need to be adapted.For samples with unknown abundance,repeated adjustment of droplet size is not allowed.In this study,a one-step double-volume droplet generation method was proposed,and a double-volume droplet microfluidic chip was developed to verify the quantitative detection capability of the chip using a duck-derived kit.The results showed that the droplets with different volumes had different quantitative capabilities.Large-volume droplets had higher reliability for low-abundance sample concentrations,while small-volume droplets had advantages in detecting high-abundance sample concentrations.The double-volume droplets produced by the double-volume droplet microfluidic chip proposed in this study greatly improved the reliability of quantitative capabilities,and had broad application prospects in detection of precious nucleic acid samples with unknown abundance in the field of microfluidic PCR.关键词
双体积液滴/数字聚合酶链式反应/绝对定量/动态检测范围Key words
Double-volume droplets/Digital polymerase chain reaction/Absolute quantification/Dynamic detection range引用本文复制引用
李姗姗,曹云亮,赵雪毅,李军委..双体积液滴对数字液滴PCR定量检测能力提升的研究[J].分析化学,2025,53(7):1138-1145,8.基金项目
国家自然科学基金项目(No.52475590)、天津市科技支撑重点项目(No.23YFZCSN00370)、河北省自然科学基金项目(No.F2023202049)和河北省省级科技计划项目(No.22567601H)资助. Supported by the National Natural Science Foundation of China(No.52475590),the Key Project of Science and Technology Support Program of Tianjin(No.23YFZCSN00370),the Hebei Natural Science Foundation for Distinguished Young Scholars(No.F2023202049),and the Science&Technology Program of Hebei Province(No.22567601H). （No.52475590）
