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1、·566·胃肠病学和肝病学杂志2008年7月第17卷第7期ChinJGastroenterolHepatol,Jul2008,Vol.17,No.7细菌16SrDNA荧光定量PCR法分析溃疡性结肠炎患者肠道菌群变化白鹏,吕愈敏,顾芳北京大学第三医院消化科,北京100083【摘要】目的应用实时荧光定量PCR技术对溃疡性结肠炎(ulcerativecolitis,UC)患者的粪便菌群进行定量分析。方法研究对象包括30名UC活动期、30名UC缓解期患者及22名正常对照,收集其粪便标本。根据细菌的16SrDNA序列设计双歧杆菌属、乳酸杆菌属、肠球菌属和大肠杆菌的特异性引物。待测粪便标本提取细菌
2、基因组DNA,进行实时荧光定量PCR反应测定16SrDNA拷贝数,分析不同细菌的数量。结果UC活动期患者与正常对照相比,粪便中双歧杆菌(8.45±0.92vs9.10±0.78)、乳酸杆菌(8111±0.94vs8.69±0.55)数量明显减少(P<0.05),大肠杆菌(9.88±1.34vs9.71±0.92)和肠球菌(7.74±0.63vs7.61±0149)无明显变化(P>0.05);而UC缓解期患者粪便中4种细菌数量(双歧杆菌9.15±0.81,乳酸杆菌8151±0.80,大肠杆菌9.54±1.64,肠球菌7.90±0.46)均与正常对照无明显差异(P>0.05)。结论UC活动
3、期患者粪便双歧杆菌、乳酸杆菌数量较正常对照明显减少,而UC缓解期患者粪便菌群与正常对照无明显差异。提示肠道菌群失调可能是具有UC遗传易感性个体发病的触发因素。【关键词】溃疡性结肠炎;肠道菌群;16SrDNA分析;实时定量PCR中图分类号:R575.2文献标识码:A文章编号:1006-5709(2008)07-0566-06收稿日期:200820222816SrDNA2basedfluorescentreal2timePCRanalysisofthevariationoffecalmicrobiotainpa2tientswithulcerativecolitisBAIPeng,LVYu
4、min,GUFangDepartmentofGastroenterology,PekingUniversityThirdHospital,Beijing100083,China【Abstract】ObjectiveToanalyzethefecalmicrobiotaofUCpatientssubjectsusingfluorescentreal2timePCR.MethodsFreshfecalsampleswereobtainedfrompatientswithactiveUC(n=30),UCinremission(n=30),andhealthyvolunteers(n=22)
5、.Asetof16SrDNA2targetedgroup2orspecies2specificprimersforBifidobacteriumspp.,Lactobacillusgroup,Enterococcusspp.andEscherichiacoliweredesigned.16SrDNAcopynumbersofbacterialgenomeDNAextractedfromfecalsampleswerequantifiedbyreal2timePCRtoanalyzebacterialamounts.ResultsThelevelsofBifidobacteriumspp
6、.(8.45±0.92vs9.10±0.78)andLactobacillusgroup(8.11±0.94vs8.69±0.55)inactiveUCsamplesweresignificantlylowerthanthoseofhealthycontrols(P<0.05),whereasEnterococcusspp.(7.74±0.63vs7.61±0.49)andEscherichiacoli(9.88±1.34vs9.71±0.92)levelsdidnotshowsignificantdifference(P>0.05).However,allfourfecalbacte
7、riaquantitiesofUCpatientsinremission(9.15±0.81,8.51±0.80,7.90±0.46,9.54±1.64,respectively)weresimilartothoseofhealthycontrols(P>0.05).ConclusionThefecalfloracompositionofactiveUCpatientshadasignificantdecreaseinBifidobacteri
