CAJ | 학술논문

目的探讨慢性HBV感染者肠道真菌菌群的分子生态结构变化特点.方法采用真菌18S rDNA通用引物对乙肝肝硬化患者、慢性乙型肝炎患者、HBV携带者和健康志愿者4组研究对象的粪便标本DNA进行PCR扩增,扩增产物进行18S rRNA基因克隆,构建真菌18S rDNA克隆文库,利用限制性片段长度多态性(RFLP)技术筛选阳性克隆子并进行测序,绘制系统发育树,获取各组研究对象肠道真菌菌群结构特征.结果所有阳性克隆子经过酶切分析和测序,共获得29个操作分类单元(OTUs),归属于3个真菌类群:接合菌纲(3.4%)、子蓑菌纲(82.8%)和担子菌纲(13.8%),其中主要优势菌属为念珠菌属(Candida spp.)、未能培养真菌(uncultured fungus)、酵母菌属(Saccharomyces spp.),分别占克隆文库的29.2%、15.9%、15.0%.乙肝肝硬化患者组、慢性乙型肝炎患者组、HBV携带者组和健康志愿者组的肠道真菌菌群分别存在有20、16、12、14个OTUs.结论人类肠道中存在较为丰富的真菌类群,慢性HBV感染者肠道真菌菌群分子生态结构发生明显改变,提示肠道真菌菌群生态结构改变与乙肝发展历程相关.
목적 탐토만성HBV감염자장도진균균군적분자생태결구변화특점.방법 채용진균18S rDNA통용인물대을간간경화환자、만성을형간염환자、HBV휴대자화건강지원자4조연구대상적분편표본DNA진행PCR확증,확증산물진행18S rRNA기인극륭,구건진균18S rDNA극륭문고,이용한제성편단장도다태성(RFLP)기술사선양성극륭자병진행측서,회제계통발육수,획취각조연구대상장도진균균군결구특정.결과 소유양성극륭자경과매절분석화측서,공획득29개조작분류단원(OTUs),귀속우3개진균류군:접합균강(3.4%)、자사균강(82.8%)화담자균강(13.8%),기중주요우세균속위념주균속(Candida spp.)、미능배양진균(uncultured fungus)、효모균속(Saccharomyces spp.),분별점극륭문고적29.2%、15.9%、15.0%.을간간경화환자조、만성을형간염환자조、HBV휴대자조화건강지원자조적장도진균균군분별존재유20、16、12、14개OTUs.결론 인류장도중존재교위봉부적진균류군,만성HBV감염자장도진균균군분자생태결구발생명현개변,제시장도진균균군생태결구개변여을간발전역정상관.
Objective To investigate the ecological structure of intestinal fungal flora of patients with chronic hepatitis B infection. Methods HBV-liver cirrhosis patients, chronic hepatitis B patients, HBV carriers, and healthy volunteers were selected as research subjects to extract the total DNA from stool of each subject, 18S rRNA genes of intestinal fungi was cloned and sequence was analyzed for the construction of 18S rDNA clone libraries and phylogenetic tree, diversity and structural characteristics of intestinal fungal flora in each group was then analyzed. Results A total of 29 operational taxonomic units (OTUs) of intestinal fungal flora in all research subjects was acquired. All positive clones belonged to three fungal taxa: Zygomycetes (3.4%) , Ascomycetes (82. 8%) and Basidiomycetes (13. 8%) . The dominant fungal community was Candida spp. , uncultured fungus and Saccharomyces spp. , which accounted for 29. 2% , 15. 9%, 15.0% respectively. The OTUs which belonged to HBV-liver cirrhosis patients, chronic hepatitis B patients, HBV carriers, and healthy volunteers was 20,16,12,14 respectively. Conclusion There is an abundant fungal flora in the human intestine. Ecological structure of intestinal fungal flora appears to be various in patients with chronic hepatitis B infection, which indicate the close relationship between the alteration of ecological structure of fungal flora and the stage of chronic hepatitis B infection.