腹泻患者肠道菌群数量变化及ERIC-PCR指纹图谱分析

Analysis of the changes of intestinal microflora in diarrhea patients by fluorescent quantitative real-time PCR and ERIC-PCR

目的了解粪检有白细胞的腹泻患者与健康者肠道菌群变化及指纹图谱差异。方法提取解放军第252医院临床门诊30名健康成人和30例腹泻成人的粪便总DNA为模板,分为对照组和腹泻组。荧光定量法测定肠道内双歧杆菌、乳酸杆菌、大肠埃希菌、肠球菌的菌群数量,ERIC-PCR指纹图谱法检查各组肠道菌群组成多样性差异。结果两组肠道菌群数量:对照组粪便中双歧杆菌为(8.85±0.57)拷贝/g,乳酸杆菌为(8.36±0.45)拷贝/g;腹泻组粪便中双歧杆菌为(8.28±0.68)拷贝/g,乳酸杆菌为(7.79±0.39)拷贝/g;与对照组比较,腹泻组中该两种细菌数量明显减少(P<0.05)。大肠埃希菌和肠球菌数量在两组之间差异无统计学意义(P>0.05)。腹泻患者肠道菌群ERIC-PCR指纹图谱条带明显减少。结论腹泻患者的肠道菌群结构发生了改变,菌群数量明显减少。

Objective To analyze the diversity variation and the fingerprints difference of intestinal microflora between diarrhea patients with the fecal leukocytes and health people. Methods 30 cases of normal people and 30 cases of diarrhea patients in No.252 Hospital of PLA were selected, whose total fecal DNA were used as model, were divided into control group and diarrhea group. Intestinal Bifidobacterium, Lactobacillus, Escherichia Coli, Enterococci bacteria number were detected by quantitative fluorescence method, the difference of intestinal flora diversity in the two groups was detected by ERIC-PCR fingerprint method. Results The quantities of intestinal microflora in the control group and di-arrhea group：Bifidobacterium spp.[（8.85±0.57） copies/g vs（8.28±0.68） copies/g];Lactobacillus spp.[（8.36±0.45） copies/g vs（7.79±0.39） copies/g], there were significant differences （P〈0.05）. There was no difference about the Escherichia Coli and Enterococci in the two groups （P〉0.05）. The bands of ERIC-PCR fingerprints in diarrhea group were obviously declined. Conclusion The constitution of the intestinal microflora is changed in diarrhea patients and the quantity of intestinal flora is significant declined.