Effects of diverse materials-based methods on DNA extraction for Clostridium difficle from stool samples
Effects of diverse materials-based methods on DNA extraction for Clostridium difficle from stool samples
复制标题
不同材料方法对粪便样本中艰难梭菌 DNA 提取的影响
DOI：
10.1166/mex.2019.1520
复制
发表时间：
2019-08
影响因子：
0.7
通讯作者：
Chen Hui
中科院分区：
文献类型：
--
作者：
Xiao Ziqi;Yang Gaojian;Yan Deng;Li Song;Chen Zhu;Li Wen;Wu Yanqi;Chen Hui
医院感染，包括艰难梭菌感染（CDI）及其死亡率在过去几十年中有所增加。尽管新兴的分子诊断技术具有快速、准确的结果，但从粪便样本中提取核酸仍然是第一个
Nosocomial infections, including Clostridium difficile infection (CDI), and their fatality rates have increased in the past few decades. Despite emerging molecular diagnostic technologies with rapid, accurate outcomes, nucleic acid extraction from stool samples remains the first
limiting step before downstream applications. Commercial nucleic acid extraction kits greatly decrease labor and time requirements, and also provide nucleic acid preparations with higher quality and purity for enzyme digestion analysis or genotyping. The magnetic bead based technique is a
novel method compared with the conventional spin-column method, and currently has widespread use in nucleic acid extraction. We evaluated five DNA extraction kits with magnetic beads using materials with various properties (particle size, concentration of magnetic beads, grinding beads) and
reagents (proteinase K, lysozyme, isopropanol, and absolute ethanol) to determine the cost, hands-on time, number of essential operations, and quality and purity of the DNA preparations, compared with those obtained using the QIAamp Fast DNA Stool Mini Kit. The six DNA extraction kits yielded
A260/280 ratios ranging from 0.85 to 1.9 (average 1.57), and concentrations from 3.70 to 108.09 ng/μL (average 34.64 ng/μL). All the DNA samples had acceptable downstream application effects, except for those obtained using the TIANGEN Magnetic Soil and
Stool DNA Kit. However, gel electrophoresis analysis of the DNA samples resulted in a light strip on the gel, indicating that the proteinaceous contaminant may not have been removed completely. A rapid and accurate molecular diagnostic technique could allow for more suitable treatment and
prognosis outcomes for inpatients, depending, in large part, on the quality and purity of DNA preparations, which are frequently neglected. Our study focused on the quality of commercial kits with a primary focus on the treatment of stool samples and molecular diagnostic applications.