研究人类肠道微生物组的分析方法进展。

Journal of biological methods Pub Date : 2024-11-18 eCollection Date: 2025-01-01 DOI:10.14440/jbm.2024.0050
Gijsbert J Jansen, Gerard P Schouten, Marit Wiersma
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引用次数: 0

摘要

背景:人类肠道微生物群是一个复杂的微生物生态系统,在维持人类健康中起着至关重要的作用。其组成的扰动与广泛的健康状况有关。分析技术:研究人员采用各种技术来研究肠道微生物群,每种技术都有自己的优点和局限性。聚合酶链反应(Polymerase chain reaction, PCR)是一种高度敏感的检测方法,但它依赖于DNA提取的质量。下一代测序(NGS)功能强大,但成本高昂,需要大量的数据分析。此外,NGS结果的准确性也在很大程度上取决于DNA提取过程的质量。培养方法虽然有用,但有偏见且耗时。荧光原位杂交(FISH)在可视化特定微生物种群方面表现出色,是唯一能够提供原位信息的方法。然而,直到最近,FISH在很大程度上依赖于人类对数字显微照片的解释,限制了它在高通量策略中的应用。此外,FISH的灵敏度受可见细胞数量的限制。结论:了解这些方法的优缺点对于在微生物组研究中得出可靠的结论至关重要。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Advancements in analytical methods for studying the human gut microbiome.

Background: The human gut microbiome, a complex ecosystem of microorganisms, plays a crucial role in maintaining human health. Perturbations in its composition are linked to a wide range of health conditions.

Analytical techniques: Researchers employ various techniques to study the gut microbiome, each having its own strengths and limitations. Polymerase chain reaction (PCR) is highly sensitive but dependent on the quality of DNA extraction. Next-generation sequencing (NGS) is powerful but can be costly and requires extensive data analysis. Furthermore, the accuracy of NGS results also depends heavily on the quality of the DNA extraction process. Culture methods, while useful, are biased and time-consuming. Fluorescence in situ hybridization (FISH) excels in visualizing specific microbial populations and is the only method capable of providing in situ information. However, until recently, FISH was heavily reliant on human interpretation of digital photomicrographs, limiting its application in high-throughput strategies. Additionally, the sensitivity of FISH is restricted by the number of cells visualized.

Conclusion: Understanding the strengths and weaknesses of these methods is essential for drawing robust conclusions in microbiome research.

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