中国循证儿科杂志
中國循證兒科雜誌
중국순증인과잡지
CHINESE JOURNAL OF EVIDENCE-BASED PEDIATRICS
2014年
5期
359-364
,共6页
陈娜%杨毅%张澜%方剑火%郎继东%曹云%田埂
陳娜%楊毅%張瀾%方劍火%郎繼東%曹雲%田埂
진나%양의%장란%방검화%랑계동%조운%전경
早产儿%肠道菌群%败血症%高通量测序
早產兒%腸道菌群%敗血癥%高通量測序
조산인%장도균군%패혈증%고통량측서
preterm infants%Gut microbiota%Sepsis%High-throughput sequence
目的：初步探讨NICU早产儿出生后肠道菌群变化特征及其与败血症的关系。方法以复旦大学附属儿科医院NICU住院的早产儿为研究对象，于出生后第1天采集胎粪，之后于每周龄时或评估败血症时采集粪便样本，直至出院或生后8周。采用 Illumina 高通量测序技术对粪便样本中所有细菌的16S rRNA-V3区进行 DNA 测序，应用MG-RAST V3.3.6分析和统计样品序列数目、操作分类单元（ OTU）数量，分析肠道菌群物种丰度和分布，并进行聚类分析。结果3例早产儿共采集生后1、7、14和21 d的12份粪便样本，其中5份样本pCR扩增失败，7份样本（例1生后14、21 d；例2生后7、21 d；例3生后7、14、21 d）DNA测序成功进入分析。3例早产儿平均胎龄为（31.3±0.8）周，平均出生体重为（1540±144）g。3例均生后应用抗生素，其中例1母亲有产前抗生素暴露史；例2在住院过程中发生全身炎症反应综合征。①7份样本稀疏曲线表明测序深度充分。物种多样性分析显示OTU值为381~608，微生物丰度较高，且与日龄呈正相关，其中例2生后7 d样本肠道菌群多样性最低；②7份样本共检测到18个菌门，均以放线菌门、拟杆菌门、厚壁菌门和变形菌门为优势菌门；变形菌门在例1生后14和21 d样本分别占97.52%和49.11%，放线菌门在例2生后7 d样本占99.46%；③共检测到172个科，其中63科为7份样本共有；相对丰度≥1%的科共检测到10个，例2生后7 d样本棒状杆菌科占97.90%，21 d样本中葡萄球菌科占27.16%；④聚类分析显示，同一研究对象不同时点肠道微生物相似性较高。结论早产儿产前抗生素暴露及出生后早期抗生素暴露可能会显著降低肠道微生物的多样性，影响正常菌群的定植。发展为败血症的早产儿生后肠道微生物多样性较低，以致病菌占优势的肠道微生物区可能与败血症的发生相关。
目的：初步探討NICU早產兒齣生後腸道菌群變化特徵及其與敗血癥的關繫。方法以複旦大學附屬兒科醫院NICU住院的早產兒為研究對象，于齣生後第1天採集胎糞，之後于每週齡時或評估敗血癥時採集糞便樣本，直至齣院或生後8週。採用 Illumina 高通量測序技術對糞便樣本中所有細菌的16S rRNA-V3區進行 DNA 測序，應用MG-RAST V3.3.6分析和統計樣品序列數目、操作分類單元（ OTU）數量，分析腸道菌群物種豐度和分佈，併進行聚類分析。結果3例早產兒共採集生後1、7、14和21 d的12份糞便樣本，其中5份樣本pCR擴增失敗，7份樣本（例1生後14、21 d；例2生後7、21 d；例3生後7、14、21 d）DNA測序成功進入分析。3例早產兒平均胎齡為（31.3±0.8）週，平均齣生體重為（1540±144）g。3例均生後應用抗生素，其中例1母親有產前抗生素暴露史；例2在住院過程中髮生全身炎癥反應綜閤徵。①7份樣本稀疏麯線錶明測序深度充分。物種多樣性分析顯示OTU值為381~608，微生物豐度較高，且與日齡呈正相關，其中例2生後7 d樣本腸道菌群多樣性最低；②7份樣本共檢測到18箇菌門，均以放線菌門、擬桿菌門、厚壁菌門和變形菌門為優勢菌門；變形菌門在例1生後14和21 d樣本分彆佔97.52%和49.11%，放線菌門在例2生後7 d樣本佔99.46%；③共檢測到172箇科，其中63科為7份樣本共有；相對豐度≥1%的科共檢測到10箇，例2生後7 d樣本棒狀桿菌科佔97.90%，21 d樣本中葡萄毬菌科佔27.16%；④聚類分析顯示，同一研究對象不同時點腸道微生物相似性較高。結論早產兒產前抗生素暴露及齣生後早期抗生素暴露可能會顯著降低腸道微生物的多樣性，影響正常菌群的定植。髮展為敗血癥的早產兒生後腸道微生物多樣性較低，以緻病菌佔優勢的腸道微生物區可能與敗血癥的髮生相關。
목적：초보탐토NICU조산인출생후장도균군변화특정급기여패혈증적관계。방법이복단대학부속인과의원NICU주원적조산인위연구대상，우출생후제1천채집태분，지후우매주령시혹평고패혈증시채집분편양본，직지출원혹생후8주。채용 Illumina 고통량측서기술대분편양본중소유세균적16S rRNA-V3구진행 DNA 측서，응용MG-RAST V3.3.6분석화통계양품서렬수목、조작분류단원（ OTU）수량，분석장도균군물충봉도화분포，병진행취류분석。결과3례조산인공채집생후1、7、14화21 d적12빈분편양본，기중5빈양본pCR확증실패，7빈양본（례1생후14、21 d；례2생후7、21 d；례3생후7、14、21 d）DNA측서성공진입분석。3례조산인평균태령위（31.3±0.8）주，평균출생체중위（1540±144）g。3례균생후응용항생소，기중례1모친유산전항생소폭로사；례2재주원과정중발생전신염증반응종합정。①7빈양본희소곡선표명측서심도충분。물충다양성분석현시OTU치위381~608，미생물봉도교고，차여일령정정상관，기중례2생후7 d양본장도균군다양성최저；②7빈양본공검측도18개균문，균이방선균문、의간균문、후벽균문화변형균문위우세균문；변형균문재례1생후14화21 d양본분별점97.52%화49.11%，방선균문재례2생후7 d양본점99.46%；③공검측도172개과，기중63과위7빈양본공유；상대봉도≥1%적과공검측도10개，례2생후7 d양본봉상간균과점97.90%，21 d양본중포도구균과점27.16%；④취류분석현시，동일연구대상불동시점장도미생물상사성교고。결론조산인산전항생소폭로급출생후조기항생소폭로가능회현저강저장도미생물적다양성，영향정상균군적정식。발전위패혈증적조산인생후장도미생물다양성교저，이치병균점우세적장도미생물구가능여패혈증적발생상관。
Objective To explore changes of postnatal gut microbiota in preterm infants in NICU and its association with neonatal sepsis. Methods preterm infants hospitalized in NICU of Childrenˊs Hospital of Fudan University were enrolled in the study. Fecal samples were collected at day 1,7,14 and 21 after birth,respectively. Illumina high-throughput sequencing techno-logy was used to sequence 16S rRNA-V3 hypervariable region of all microbes in 7 fecal samples of 3 recruited subjects. MG-RAST V3. 3. 6 was used to analyze and calculate the numbers of sequences and operational taxonomic units(OTUs)for each sample,then the species abundance and distribution were analyzed and followed by cluster analysis. Results Seven samples had been analyzed. Three subjects had an average gestational age of(31. 3 ± 0. 8)weeks and an average birth weight of(1 540 ± 144)g. They all received antibiotic administration after birth. Maternal antibiotic exposure occured in 1 case. Another one case developed SIRS during hospitalization. ①The rarefaction curves showed that adequate sequencing depth was achieved. Analysis of species abundance showed that the OTUs number ranged from 381 to 608 indicating high microbial diversity. Meanwhile,the microbial diversity had a positive correlation with the postnatal age. ②Eighteen phyla were detected from all samples. Actinobacteria, Bacteriodetes,Firmicutes and Proteobacteria were predominant in all samples. However,the dominant phylum of case 1 belonged to Proteobacteria,with a percentage of 97. 5%(day 14)and 49. 1%(day 21),while Actinobacteria predominated in day-7 sample of case 2,with a percentage of 99. 5%. ③172 families were detected altogether,63 of them were detected in all samples. Ten families were in higher relative abundance. In case 2,Corynebacteria occupied 97. 9% in day-7 sample,and Staphylococcaceae occupied a higher proportion in day-21 sample(27. 2%)than the other two samples. ④The cluster analysis showed high similarity of intestinal microflora in different time points of one subject. Conclusion Gut microbial colonization and development in preterm infants in NICU were altered by various factors. prolonged broad-spectrum antibiotics at prenatal and early postnatal age might profoundly decrease microbial diversity and affect microbial colonisation. Microbiota was less diverse from birth in infants who developed sepsis. There may be a microbiome predominanted of pathogens that may be associated with sepsis in preterm infants.
