Evaluation of freeze-drying for quantification of the microbiome and metabolome in neonatal faecal samples

Wei Shen , Di Wu , Wen Qiu , Xinzhu Yi
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引用次数: 3

Abstract

Appropriate sampling and storage processes are crucial for the accurate determination of the gut microbiome and metabolome. The present study evaluated the effect of the freeze-drying process on meconium and regular faecal samples of newborn babies. Triplicate samples were freeze-dried for 6 or 12 ​h and compared to samples that were only frozen. Microbiome analysis using 16 ​S rRNA sequencing showed that the microbial composition was similar between samples with freeze-drying and those without freeze-drying in general, while the determination efficiency of Bifidobacterium was significantly increased after freeze-drying for either time. A total of 300 commonly observed gut metabolites were quantified and classified into 13 categories, including amino acids, bile acids, carbohydrates, carnitines, fatty acids and so on. The compositions of most metabolites were consistently preserved before or after freeze-drying for the two time protocols. Moreover, the sensitivity of detection was significantly improved after freeze-drying, even for volatile compounds such as short-chain fatty acids. Accordingly, freeze-drying for 6 ​h is recommended for the pretreatment of bulk meconium and regular faecal samples for the determination of gut microbiota.

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冷冻干燥定量新生儿粪便样品中微生物组和代谢组的评价
适当的采样和储存过程对于准确测定肠道微生物组和代谢组至关重要。本研究评估了冷冻干燥过程对新生儿胎便和正常粪便样本的影响。将三份样品冷冻干燥6或12小时,并与仅冷冻的样品进行比较。采用16s rRNA测序对样品进行微生物组分析,结果表明,冷冻干燥样品的微生物组成与未冷冻干燥样品的微生物组成基本相似,而双歧杆菌的测定效率在两次冷冻干燥后均有显著提高。对300种常见的肠道代谢物进行量化,并将其分为13类,包括氨基酸、胆汁酸、碳水化合物、肉碱、脂肪酸等。在两种时间方案中,大多数代谢物的组成在冷冻干燥前后保持一致。此外,冷冻干燥后的检测灵敏度显著提高,即使对挥发性化合物如短链脂肪酸也是如此。因此,建议冷冻干燥6小时预处理散装胎便和常规粪便样品,以测定肠道微生物群。
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来源期刊
Medicine in Microecology
Medicine in Microecology Medicine-Gastroenterology
CiteScore
5.60
自引率
0.00%
发文量
16
审稿时长
76 days
期刊最新文献
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