CAJ | 학술논문

目的：探讨16S rDNA测序技术在新生儿、婴儿肠道微生态研究中的应用。方法于生后3天、1月、6月、1岁时收集2例健康婴儿粪便标本共8份，提取细菌总DNA ，以Illumina Hiseq 2000为测序平台，采用新一代高通量16S rDNA宏基因组测序技术对V6可变区测序，并进行生物信息分析（物种分类和丰度分析；多样性分析）。结果8份样品共产生原始测序数据为1027．47 Mbp ，Unique tags序列数量均值为58630，OTU数量63～209；优势菌门为Proteobacteria和Firmicutes ；在科水平，＞1％的物种1个月之内2～4种，6月后达7～10种；1号婴儿一直以Enterobacteriaceae占优势，2号婴儿优势菌群包括 Enterobacteriaceae、Lachnospiraceae、Streptococcaceae和Bacteroidaceae；4个时间点的npShannon和Simpson指数分别为1．17、1．29、2．16、2．51和0．43、0．40、0．26、0．14。结论16S rDNA测序技术能满足新生儿、婴儿肠道微生态研究需求；新生儿、婴儿粪便中含丰富细菌基因组；细菌物种丰度及分类存在个体差异；从出生到1岁，婴儿肠道菌群结构趋向复杂和多样。
목적：탐토16S rDNA측서기술재신생인、영인장도미생태연구중적응용。방법우생후3천、1월、6월、1세시수집2례건강영인분편표본공8빈，제취세균총DNA ，이Illumina Hiseq 2000위측서평태，채용신일대고통량16S rDNA굉기인조측서기술대V6가변구측서，병진행생물신식분석（물충분류화봉도분석；다양성분석）。결과8빈양품공산생원시측서수거위1027．47 Mbp ，Unique tags서렬수량균치위58630，OTU수량63～209；우세균문위Proteobacteria화Firmicutes ；재과수평，＞1％적물충1개월지내2～4충，6월후체7～10충；1호영인일직이Enterobacteriaceae점우세，2호영인우세균군포괄 Enterobacteriaceae、Lachnospiraceae、Streptococcaceae화Bacteroidaceae；4개시간점적npShannon화Simpson지수분별위1．17、1．29、2．16、2．51화0．43、0．40、0．26、0．14。결론16S rDNA측서기술능만족신생인、영인장도미생태연구수구；신생인、영인분편중함봉부세균기인조；세균물충봉도급분류존재개체차이；종출생도1세，영인장도균군결구추향복잡화다양。
Objective To investigate the application of 16S rDNA sequencing technique in studying intestinal microecology of neonates and infants .Methods Eight fecal samples were collected on 3 d ,1 month ,6 months and 1 year in 2 healthy infants .Total bacterial DNAs were extracted and submitted the high throughout 16S rDNA sequen‐cing on the V6 viable region by using Illumia genome analyzer Hiseq 2000 (101 bp pair‐end sequencing strategy) .The 16S rDNA tags and operational taxonomic units (OTU) were then obtained from the sequences .The bioinformatic a‐nalysis including analysis of taxonomy ,abundance and alpha diversity were performed .Results Total 1 027 .47 Mbp raw data were produced .The mean unique tags number was 58630 .The OTU numbers ranged from 36 to 308 .The bacterial families more than 1% were increased from 2-4 species per sample before 1 month to 7‐10 species after 6 months .Enterobacteriaceae was always the predominant family in No .1 infant through the first year ,while in No .2 infant the predominant groups included Enterobacteriaceae ,Lachnospiraceae ,Streptococcacea and Bacteroidaceae .The mean npShannon and Simpson indexes at 4 time points were 1 .17 ,1 .29 ,2 .16 ,2 .51 and 0 .43 ,0 .40 ,0 .26 ,0 .14 re‐spectively .Conclusion 16S rDNA sequencing could meet the need of studying intestinal microecology in neonates and infants ;neonatal and infantile feces contains abounding bacterial genomes ;the individual differences exist in intestinal bacterial abundance and composition;the structure of gut bacterial floras trend to complexity and diversity from birth to 1 year old .