两种方法定量分析肝硬化患者肠道菌群改变的比较

Comparison of two methods applied in the quantitative analysis of gut flora alteration in patients with liver cirrhosis

目的观察肝硬化患者肠道菌群改变,探讨传统培养法与实时定量PCR检测结果的一致性。方法纳入73名肝硬化患者及55名健康人,对其粪便细菌进行传统培养及定量PCR,比较两种方法检测结果的一致性。结果细菌培养结果显示,与正常对照组比较,肝硬化患者肠杆菌、肠球菌、葡萄球菌、酵母菌数量显著增加(P分别为0.004、0.002、0.044、0.026),拟杆菌、乳酸杆菌、双歧杆菌、梭菌数量显著降低(P分别为0.029、0.046、0.045、0.045)。定量PCR结果显示,肝硬化组肠杆菌、肠球菌数量显著增高(P分别<0.01、0.012),乳酸杆菌、双歧杆菌、拟杆菌及梭菌数量显著降低(P均为<0.01)。结论肝硬化患者存在不同程度肠道菌群失衡,可能影响病情发展。传统细菌培养与荧光定量PCR的结果具有良好的一致性,但定量PCR更精确、灵敏、省时、省力。

Aims The objective of our research is to explore the consistency between traditional culture and real-time quantitative PCR（qPCR） through testing fecal microbiota in patients with liver cirrhosis. Methods 73 patients with liver cirrhosis and 55 healthy controls were enrolled. Their fecal bacteria were detected by traditional culture and qPCR, and the results from two different methods were compared. Results To compare with healthy controls, culture results showed that the lever of Escherichia coli, Enterococcus, staphylococcus and yeast was significantly enriched（P = 0.004, 0.002, 0.044, 0.026, respectively）, Bacteroides, LactobaciUus, Bifidobacterium and Clostridium were significantly reduced in patient group（P = 0.029, 0.046, 0.045, 0.045, respectively）, qPCR results suggested that the amount of Escherichia coli and Enterococcus were statistically increased（P 〈 0.01, 0.012, respectively）, Lactobacillus, Bifidobacterium, Bacteroides and Clostridium were statistically decreased（all P 〈 0.01）. Conclusion Fecal microbial communities are imbalanced in cirrhotic pa- tients compared with healthy individuals, which may affect the prognosis. The results of traditional culture and qPCR are consistent, but the latter with the more accurate and sensitive, time saving, energy saving.