实时荧光定量PCR在测定艾滋病患者肠道菌群量变化中的应用及意义

目的:应用实时荧光定量PCR观察艾滋病患者肠道菌群量的变化,研究艾滋病对肠道微生态的影响及其在发病中的作用。方法:分别设计双歧杆菌属、乳酸杆菌属、大肠杆菌、粪肠球菌及屎肠球菌的特异性引物。收集艾滋病患者粪便标本30份及正常对照标本30份,提取细菌基因组DNA,应用实时荧光定量PCR反应测定5种细菌的数量。结果:艾滋病患者组双歧杆菌属、乳酸杆菌属的数量较正常对照组明显减少;大肠杆菌、粪肠球菌、屎肠球菌数量明显增多,差异有统计学意义(P<0.05)。结论:艾滋病患者肠道微生态发生了明显变化,提示艾滋病患者肠道微生态紊乱。

Real-time fluorescence quantitative PCR analysis of intestinal flora in patients with acquired immune deficiency syndrome

Objective To analyze the intestinal microflora of AIDS patients by real-time fluorescence quantitative PCR(RT-PCR),which helps explore the influence of AIDS on intestinal microflora,reveal the role and significance of gut microflora in AIDS.Methods The feces of patients with AIDS(n = 30) and healthy volunteers(n = 30) were collected.A set of species-specific primers for Bifidobacterium spp,Lactobacillus group,Escherichia coli,Enterococcus faecalis and feces Enterococcus were designed..Bacterial genome DNA was extracted from fecal samples,was quantified by RT-PCR to analyze bacterial amounts.Results The level of Bifidobacterium spp.and Lactobacillus group in AIDS group were significantly lower than those of healthy control group,whereas Escherichia coli,Enterococcus faecalis and feces Enterococcus were significantly higher than those of healthy control group(P < 0.05).Conclusion The intestinal microflora of AIDS patient have obviously changed.These data underline that the intestinal microflora of AIDS patients is disorder.