Wang Chen, Feng Saisai, Xiao Yue, Pan Mingluo, Zhao Jianxin, Zhang Hao, Zhai Qixiao, Chen Wei
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China.
School of Food Science and Technology, Jiangnan University, Wuxi, Jiangsu, 214122, China.
Microb Biotechnol. 2021 Mar;14(2):577-586. doi: 10.1111/1751-7915.13651. Epub 2020 Aug 11.
Bacteroides is a bacterial genus that is known to closely interact with the host. The potential role of this genus is associated with its ecological status and distribution in the intestine. However, the current 16S V3-V4 region sequencing method can only detect the abundance of this genus, revealing a need for a novel sequencing method that can elucidate the composition of Bacteroides in the human gut microbiota. In this study, a core gene, rpsD, was selected as a template for the design of a Bacteroides-specific primer set. We used this primer set to develop a novel assay based on the Illumina MiSeq sequencing platform that enabled an accurate assessment of the Bacteroides compositions in complex samples. Known amounts of genomic DNA from 10 Bacteroides species were mixed with a complex sample and used to evaluate the performance and detection limit of our assay. The results were highly consistent with those of direct sequencing with a low Bacteroides DNA detection threshold (0.01 ng), supporting the reliability of our assay. In addition, the assay could detect all the known Bacteroides species within the faecal sample. In summary, we provide a sensitive and specific approach to determining the Bacteroides species in complex samples.
拟杆菌属是一种已知与宿主密切相互作用的细菌属。该菌属的潜在作用与其在肠道中的生态状况和分布有关。然而，当前的16S V3 - V4区域测序方法只能检测该菌属的丰度，这表明需要一种新的测序方法来阐明人类肠道微生物群中拟杆菌属的组成。在本研究中，选择核心基因rpsD作为设计拟杆菌属特异性引物组的模板。我们使用该引物组开发了一种基于Illumina MiSeq测序平台的新型检测方法，该方法能够准确评估复杂样品中拟杆菌属的组成。将已知量的来自10种拟杆菌属物种的基因组DNA与复杂样品混合，并用于评估我们检测方法的性能和检测限。结果与直接测序的结果高度一致，拟杆菌属DNA检测阈值低（0.01 ng），支持了我们检测方法的可靠性。此外，该检测方法能够检测粪便样品中所有已知的拟杆菌属物种。总之，我们提供了一种灵敏且特异的方法来确定复杂样品中的拟杆菌属物种。
