Kim Tae Gwan, Jeong So-Yeon, Cho Kyung-Suk
Department of Environmental Science and Engineering, Ewha Womans University, 11-1 Daehyun-dong, Seodaemun-gu, Seoul 120-750, Republic of Korea.
Biotechnol Rep (Amst). 2014 Jul 5;4:1-4. doi: 10.1016/j.btre.2014.06.010. eCollection 2014 Dec.
Two different quantitative PCR platforms, droplet digital PCR (dd-PCR) and quantitative real-time PCR (qPCR), were compared in a -based methanogen community assay that quantifies ten methanogen sub-groups. Both technologies exhibited similar PCR efficiencies over at least four orders of magnitude and the same lower limits of detection (8 copies μL-DNA extract). The -based methanogen communities in three full-scale anaerobic digesters were examined using the two technologies. dd-PCR detected seven groups from the digesters, while qPCR did five groups, indicating that dd-PCR is more sensitive for DNA quantification. Linear regression showed quantitative agreements between both of the technologies ( = 0.59-0.98) in the five groups that were concurrently detected. Principal component analysis from the two datasets consistently indicated a substantial difference in the community composition among the digesters and revealed similar levels of differentiation among the communities. The combined results suggest that dd-PCR is more promising for examining methanogenic archaeal communities in biotechnological processes.
在一项基于定量分析十种产甲烷菌亚群的产甲烷菌群落检测中，对两种不同的定量PCR平台——液滴数字PCR（dd-PCR）和定量实时PCR（qPCR）进行了比较。两种技术在至少四个数量级上均表现出相似的PCR效率，且检测下限相同（8个拷贝/微升DNA提取物）。使用这两种技术对三个全尺寸厌氧消化器中基于[具体内容缺失]的产甲烷菌群落进行了检测。dd-PCR从消化器中检测到七组，而qPCR检测到五组，表明dd-PCR在DNA定量方面更敏感。线性回归显示，在同时检测的五组中，两种技术之间存在定量一致性（[相关系数缺失] = 0.59 - 0.98）。来自两个数据集的主成分分析一致表明，消化器之间的群落组成存在显著差异，并揭示了群落之间相似的分化水平。综合结果表明，dd-PCR在检测生物技术过程中产甲烷古菌群落方面更具前景。
