肾结石患者肠道内草酸降解细菌的定量检测

Quantification of Oxalate-degrading Bacteria in the Gut of Kidney Stone Patients Using Real-time PCR

为了分析肠道内主要草酸降解细菌与肾结石疾病的相关性,本研究采集 29 名健康对照和 30 名肾结石患者的粪便样品提取微生物基因组 DNA,采用实时荧光定量 PCR 的方法,检测产甲酸草酸杆菌Oxalobacter formigenes)、乳杆菌属细菌 (Lactobacillus)、双歧杆菌属细菌 (Bifidobacterium) 和肠杆菌科细菌(Enterobacteriaceae)这 4 种草酸降解细菌功能类群在总菌中相对含量的变化。结果显示 O. formigenes、Lact-%obacillus 和 Bifidobacterium 在肾结石病例组与健康对照组的肠道内的丰度无显著差异,而 Enterobacteriaceae在两组之间的差异显著。与健康对照组相比,Enterobacteriaceae 在肾结石病例组的丰度偏高(p=0.02);将肾结石病例组按照结石发生位置分为单发组和多发组,Enterobacteriaceae 在健康对照组与结石单发组的丰度无显著差异,但在结石多发组的丰度都显著高于健康对照组(p=0.001)与结石单发组(p=0.042)。结果提示Enterobacteriaceae 丰度可以作为一个灵敏的生物学指标,用于预测和评价宿主罹患多发肾结石的风险,有助于更深入的研究宿主和肠道菌群之间在肾结石发生发展中的相互作用,为从维持肠道微生态平衡角度进行针对性治疗提供参考。

To quantify the difference of oxalate-degrading bacteria in the gut of kidney stone patients and healthy individuals using Real-time PCR, we extracted fecal DNA from 29 healthy individuals and 30 kidney stone patients. Real-time PCR was employed to quantify the difference of Oxalobacterformigenes, Lactobacillus, Bifidobacterium and Enterobacteriaceae in the gut microbiome of healthy individuals and kidney stone patients. There was no significant difference of relative percentage of O. formigenes, Lactobacillus and Bifidobacteriura between kidney stone patients and healthy individuals, but the abundance of Enterobacteriaceae in kidney stone patients was signif- icantly higher than the abundance in healthy individuals （p=0.02）. The kidney stone patients were divided into singlegroup with unique stone location and multiple-group with multiple stone locations. There was no significant differ- ence of relative percentage of Enterobacteriaceae between single-group and healthy individuals, but the abundance of Enterobacteriaceae in multiple-group was significantly higher than both the abundance in healthy individuals （p=0.001） and the abundance in single-group （p=0.042）. The results suggested that Enterobacteriaceae abundance could be used as a sensitive biological indicator for predicting and evaluating the risk of host suffering from kidney stones occuring in multiple locations, and paved the way for further study on the interaction ofgnt microbiota and host.