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基于液滴数字PCR和定量实时PCR的产甲烷菌群落分析比较

Comparison of droplet digital PCR and quantitative real-time PCR in -based methanogen community analysis.

作者信息

Kim Tae Gwan, Jeong So-Yeon, Cho Kyung-Suk

机构信息

Department of Environmental Science and Engineering, Ewha Womans University, 11-1 Daehyun-dong, Seodaemun-gu, Seoul 120-750, Republic of Korea.

出版信息

Biotechnol Rep (Amst). 2014 Jul 5;4:1-4. doi: 10.1016/j.btre.2014.06.010. eCollection 2014 Dec.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0d35/5466119/3d8356deb329/gr1.jpg

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