Choice of 16S Ribosomal RNA Primers Impacts Male Urinary Microbiota Profiling.
Choice of 16S Ribosomal RNA Primers Impacts Male Urinary Microbiota Profiling.
复制标题
DOI：
10.3389/fcimb.2022.862338
复制
发表时间：
2022
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
--
作者：
下一代测序（NGS）技术的可及性使得能够对生活在不同栖息地的微生物群落进行分析。16S核糖体RNA（rRNA）基因测序广泛用于NGS技术的微生物群分析。由于大多数使用的NGS平台产生短读段，因此对全长16S rRNA基因进行测序是不切实际的。因此，选择哪个16S rRNA高变区进行测序在微生物群分析研究中至关重要。所有9个16S rRNA高变区都是分类学上的信息，但由于特定分支的分析性能的变化，选择理想的16S rRNA高变区将取决于所研究的栖息地的细菌组成。最近，NGS允许鉴定尿路中的微生物，并且尿微生物群已成为一个活跃的研究领域。然而，目前还没有研究评估不同的16S rRNA高变区用于男性尿液微生物群分析的性能。我们收集了男性志愿者的尿液样本，并通过对一组包含所有9个16S rRNA高变区的6个扩增子进行测序来分析他们的尿液微生物群。它们的性能的系统比较表明V1V2高变区更好地评估了男性尿液样本中常见的分类群，这表明V1V2扩增子测序更适合男性尿液微生物群分析。我们相信，我们的研究结果将有助于指导未来男性尿液微生物群研究中这一关键方法的选择。
Accessibility to next-generation sequencing (NGS) technologies has enabled the profiling of microbial communities living in distinct habitats. 16S ribosomal RNA (rRNA) gene sequencing is widely used for microbiota profiling with NGS technologies. Since most used NGS platforms generate short reads, sequencing the full-length 16S rRNA gene is impractical. Therefore, choosing which 16S rRNA hypervariable region to sequence is critical in microbiota profiling studies. All nine 16S rRNA hypervariable regions are taxonomically informative, but due to variability in profiling performance for specific clades, choosing the ideal 16S rRNA hypervariable region will depend on the bacterial composition of the habitat under study. Recently, NGS allowed the identification of microbes in the urinary tract, and urinary microbiota has become an active research area. However, there is no current study evaluating the performance of different 16S rRNA hypervariable regions for male urinary microbiota profiling. We collected urine samples from male volunteers and profiled their urinary microbiota by sequencing a panel of six amplicons encompassing all nine 16S rRNA hypervariable regions. Systematic comparisons of their performance indicate V1V2 hypervariable regions better assess the taxa commonly present in male urine samples, suggesting V1V2 amplicon sequencing is more suitable for male urinary microbiota profiling. We believe our results will be helpful to guide this crucial methodological choice in future male urinary microbiota studies.