Jansen Gijsbert J, Schouten Gerard P, Wiersma Marit
NL-Lab, Biotrack, Leeuwarden, Friesland, 8912 AP Netherlands.
J Biol Methods. 2024 Nov 18;12(1):e99010038. doi: 10.14440/jbm.2024.0050. eCollection 2025.
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.
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 hybridization (FISH) excels in visualizing specific microbial populations and is the only method capable of providing 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.
Understanding the strengths and weaknesses of these methods is essential for drawing robust conclusions in microbiome research.
人类肠道微生物群是一个复杂的微生物生态系统,在维持人类健康方面起着至关重要的作用。其组成的扰动与多种健康状况相关。
研究人员采用各种技术来研究肠道微生物群,每种技术都有其自身的优势和局限性。聚合酶链反应(PCR)高度灵敏,但依赖于DNA提取的质量。新一代测序(NGS)功能强大,但成本高昂且需要大量数据分析。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,但存在偏差且耗时。荧光杂交(FISH)在可视化特定微生物群体方面表现出色,是唯一能够提供信息的方法。然而,直到最近,FISH严重依赖于对数码显微照片的人工解读,限制了其在高通量策略中的应用。此外,FISH的灵敏度受到可视化细胞数量的限制。
了解这些方法的优缺点对于在微生物组研究中得出可靠结论至关重要。