Study on Improvement of Quantitative Capacity of Digital Droplet PCR by Double-Volume Droplets
10.19756/j.issn.0253-3820.251066
- VernacularTitle:双体积液滴对数字液滴PCR定量检测能力提升的研究
- Author:
Shan-Shan LI
1
;
Yun-Liang CAO
;
Xue-Yi ZHAO
;
Jun-Wei LI
Author Information
1. 河北工业大学机械工程学院,天津 300401;河北省机器人传感与人机融合重点实验室,天津 300130;电工装备可靠性与智能化国家重点实验室,天津 300132
- Keywords:
Double-volume droplets;
Digital polymerase chain reaction;
Absolute quantification;
Dynamic detection range
- From:
Chinese Journal of Analytical Chemistry
2025;53(7):1138-1145
- CountryChina
- Language:Chinese
-
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.
