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Abstract：
A microbial imbalance called dysbiosis leads to inflammatory bowel disease (IBD), which can include ulcerative colitis (UC). Fecal microbiota transplantation (FMT), a novel therapy, has recently been successful in treating gut dysbiosis in UC patients. For the FMT technique to be successful, the gut microbiota of both the healthy donors and UC patients must be characterized. For decades, next-generation sequencing (NGS) has been used to analyze gut microbiota. Despite the popularity of NGS, the cost and time constraints make it difficult to use in emergency services and activities related to the periodic monitoring of microbiota profile alterations. Hence, in this study, we developed a multiplex TaqMan qPCR assay (MTq-PCR) with novel probes to simultaneously determine the relative proportions of the three dominant microbial phyla in the human gut: Bacteroidetes, Firmicutes, and Proteobacteria. The relative proportions of the three phyla in fecal samples of either healthy volunteers or UC patients were similar when assessed NGS and the MTq-PCR. Thus, our MTq-PCR assay could be a practical microbiota profiling alternative for diagnosing and monitoring gut dysbiosis in UC patients during emergency situations, and it could have a role in screening stool from potential FMT donors.
摘要：
一种称为菌群失调的微生物失衡导致炎症性肠病（IBD），其中包括溃疡性结肠炎（UC）。粪便微生物移植（FMT），一种新颖的疗法,最近已成功治疗UC患者的肠道菌群失调。为了使FMT技术成功，必须对健康供体和UC患者的肠道微生物群进行表征。几十年来,下一代测序（NGS）已用于分析肠道微生物群。尽管NGS很受欢迎，成本和时间的限制使其难以用于与定期监测微生物区系变化有关的紧急服务和活动。因此，在这项研究中，我们开发了一种具有新型探针的多重TaqManqPCR测定（MTq-PCR），以同时确定人类肠道中三种优势微生物门的相对比例：拟杆菌，Firmicutes,和变形杆菌。当评估NGS和MTq-PCR时，健康志愿者或UC患者的粪便样品中三个门的相对比例相似。因此，我们的MTq-PCR分析可能是一种实用的微生物群分析替代方案，用于诊断和监测UC患者在紧急情况下的肠道菌群失调，它可能在筛查潜在FMT捐赠者的粪便中发挥作用。
