16S rDNA测序技术在婴儿肠道微生态研究中的应用  

Application of 16S rDNA sequencing technique in infantile intestinal microecological research

作　　者：马丽亚[1] 王辉林[2] 陈睿[1] 张敏[1] 黄艳[1] 卢光进[1] 

机构地区：[1]深圳市宝安区妇幼保健院新生儿科,518133 [2]深圳市宝安区妇幼保健院中心实验室,518133

出　　处：《检验医学与临床》2015年第2期166-168,共3页Laboratory Medicine and Clinic

基　　金：广东省深圳市科技计划项目(201102030)

摘　　要：目的探讨16SrDNA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16SrDNA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1 027.47 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 16SrDNA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。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 .

关 键 词：肠道菌群 测序 宏基因组 婴儿 

分 类 号：R378.16[医药卫生—病原生物学]