An ambient-temperature storage and stabilization device performs comparably to flash-frozen collection for stool metabolomics in infants.
An ambient-temperature storage and stabilization device performs comparably to flash-frozen collection for stool metabolomics in infants.
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DOI：
10.1186/s12866-021-02104-6
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发表时间：
2021-02-22
期刊：
影响因子：
4.2
通讯作者：
Hourigan SK
中科院分区：
文献类型：
--
作者：
Ramamoorthy S;Levy S;Mohamed M;Abdelghani A;Evans AM;Miller LAD;Mehta L;Moore S;Freinkman E;Hourigan SK
粪便代谢物提供了对肠道微生物组功能的重要见解。目前储存代谢组学粪便样本的黄金标准是在− 80 °C下快速冷冻，这可能不方便且昂贵。粪便的室温储存更实用，然而没有可用的方法充分保存粪便的代谢组学特征。一种新的取样试剂盒（OMNImet.GUT; DNA Genotek，Inc.）用于代谢组学的粪便环境温度储存和稳定;我们旨在测试该试剂盒与快速冷冻相比的性能。为此，从婴儿尿布收集粪便，将其分成两个等分试样：1）快速冷冻和2）在环境温度下储存在OMNImet.GUT管中3-4天。在2个不同时间点采集同一婴儿的样本，以评估代谢物随时间的变化。随后，通过液相色谱-串联质谱法（LC-MS/MS）对所有样本进行代谢组学分析。在2个时间点（32份个体样本，64份等分试样）采集16名婴儿的成对粪便样本（速冻和环境温度）。在冷冻和环境温度样品中检测到相似数量的代谢物（冷冻样品中为1126，环境温度样品中为1107，样品类型之间共有1064）。代谢物丰度在储存方法之间具有强相关性（代谢物之间的中位斯皮尔曼相关性Rs = 0.785）。系统聚类分析和主成分分析表明，样品来自同一个人在一个给定的时间点聚集紧密，无论储存方法。通过配对t检验分别比较冷冻和OMNImet. GUT的来自同一个体的重复样本。在时间点2相对于时间点1显著变化（p < 0.05）的每种生化类别中的代谢物数量在快速冷冻与环境温度储存中相似。微生物群修饰的代谢物随时间的变化在两种方法中也是一致的。在OMNImet.GUT装置中室温储存和稳定粪便产生了与快速冷冻相当的代谢组学结果，包括1）检测到的生化物质的同一性和丰度，2）受试者的不同代谢组学特征，3）可能由微生物群诱导的代谢物随时间的变化。该方法潜在地为粪便代谢组学分析提供了更方便、更便宜的家庭收集和储存选择。在线版本包含补充材料，可通过10.1186/s12866-021-02104-6获得。
Stool metabolites provide essential insights into the function of the gut microbiome. The current gold standard for storage of stool samples for metabolomics is flash-freezing at − 80 °C which can be inconvenient and expensive. Ambient temperature storage of stool is more practical, however no available methodologies adequately preserve the metabolomic profile of stool. A novel sampling kit (OMNImet.GUT; DNA Genotek, Inc.) was introduced for ambient temperature storage and stabilization of feces for metabolomics; we aimed to test the performance of this kit vs. flash-freezing. To do this stool was collected from an infant’s diaper was divided into two aliquots: 1) flash-frozen and 2) stored in an OMNImet.GUT tube at ambient temperature for 3–4 days. Samples from the same infant were collected at 2 different time points to assess metabolite changes over time. Subsequently, all samples underwent metabolomic analysis by liquid chromatography – tandem mass spectrometry (LC-MS/MS). Paired fecal samples (flash-frozen and ambient temperature) from 16 infants were collected at 2 time points (32 individual samples, 64 aliquots). Similar numbers of metabolites were detected in both the frozen and ambient temperature samples (1126 in frozen, 1107 in ambient temperature, 1064 shared between sample types). Metabolite abundances were strongly correlated between storage methods (median Spearman correlation Rs = 0.785 across metabolites). Hierarchical clustering analysis and principal component analysis showed that samples from the same individuals at a given time point clustered closely, regardless of the storage method. Repeat samples from the same individual were compared by paired t-test, separately for the frozen and OMNImet.GUT. The number of metabolites in each biochemical class that significantly changed (p < 0.05) at timepoint 2 relative to timepoint 1 was similar in flash-frozen versus ambient temperature storage. Changes in microbiota modified metabolites over time were also consistent across both methodologies. Ambient temperature storage and stabilization of stool in the OMNImet.GUT device yielded comparable metabolomic results to flash freezing in terms of 1) the identity and abundance of detected biochemicals 2) the distinct metabolomic profiles of subjects and 3) changes in metabolites over time that are plausibly microbiota-induced. This method potentially provides a more convenient, less expensive home collection and storage option for stool metabolomic analysis. The online version contains supplementary material available at 10.1186/s12866-021-02104-6.
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影响因子：
30.8
作者：
Zierer J;Jackson MA;Kastenmüller G;Mangino M;Long T;Telenti A;Mohney RP;Small KS;Bell JT;Steves CJ;Valdes AM;Spector TD;Menni C
通讯作者：
Menni C
影响因子：
4.3
作者：
Kaur, Harrisham;Bose, Chandrani;Mande, Sharmila S.
通讯作者：
Mande, Sharmila S.
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影响因子：
64.8
作者：
Wang, Zeneng;Klipfell, Elizabeth;Bennett, Brian J.;Koeth, Robert;Levison, Bruce S.;Dugar, Brandon;Feldstein, Ariel E.;Britt, Earl B.;Fu, Xiaoming;Chung, Yoon-Mi;Wu, Yuping;Schauer, Phil;Smith, Jonathan D.;Allayee, Hooman;Tang, W. H. Wilson;DiDonato, Joseph A.;Lusis, Aldons J.;Hazen, Stanley L.
通讯作者：
Hazen, Stanley L.
影响因子：
16.6
作者：
Roager HM;Licht TR
通讯作者：
Licht TR
影响因子：
2
作者：
Ford, Lisa;Kennedy, Adam D.;Toal, Douglas R.
通讯作者：
Toal, Douglas R.