摘要
目的应用PCR-变性梯度凝胶电泳(PCR-DGGE)技术动态分析分娩方式对新生儿早期肠道菌群结构的影响。方法选择32例同期出生的健康足月儿为研究对象,按分娩方式分为自然分娩组和剖宫产组(每组各16例),分别于出生后第1天(>12 h)、第3天及第5天采集粪便样本,抽提样本中细菌基因组总DNA,并对其16S rDNA V3可变区进行PCR扩增和DGGE测定,获得肠道菌群指纹图谱,进行多样性和相似性等特征分析。结果 1.新生儿出生第1天肠道菌群定植较少,生命早期肠道菌群定植具有个体特异性,并呈动态变化。2.多样性分析显示:出生第3天2组间多样性比较差异无统计学意义;第5天剖宫产组多样性指数H'和优势度指数D均显著低于自然分娩组(Pa<0.05)。3.成对相似性分析:出生第3天,剖宫产组内成对相似性系数Cs高于自然分娩组;剖宫产组内Cs值<0.30的占该组总Cs值个数的百分比(21.4%)显著低于自然分娩组(76.0%)(Pa<0.01)。第5天,2组Cs值及分布情况比较无明显差异。结论出生第3天,剖宫产个体间较自然分娩儿表现出更高的菌群结构相似性,第5天剖宫产儿肠道菌群种类及数量均落后于自然分娩儿,分娩方式影响了生命早期肠道菌群结构的建立。PCR-DGGE法在新生儿肠道菌群结构多态性及演进变化方面提供了更详尽的资料。
Objective To investigate the effects of delivery mode on the dynamic changes of early gut microflora by using polymerase chain reaction - degeneration gradient gel electrophoresis （ PCR - DGGE） method. Methods Thirty - two normal term neonates were enrolled in the study （including vaginal delivery group and caesarean section group, each group including 16 neonates） , and feces samples were col- lected on the first day（ 〉 12 h） ,3^th day and 5^th day after birth. After bacterial DNAs were extracted from the feces of the 2 groups, bacterial communities in 2 groups were examined by PCR of 16S rDNA V3 region and DGGE. After DGGE profilings were obtained,the diversity and similarity related indices were used to analyze diversity differences of the gut bacterium structure between 2 groups and the similar degrees be- tween individuals within each group. Results 1. There were few species and quantities in the intestinal microbiota during the first day of life, and the early intestinal bacterial colonization of neonates had a complex individual specificity and presented dynamic changes. 2. The diversity analysis between 2 groups showed that there were no obvious diversity differences between the 2 groups on 3th day after birth. While on 5th day after birth ,the Shannon- Weaver Diversity Index（H＇） and the simpson index（D） in caesarean section group were lower significantly than those in vaginal delivery group（Pa 〈0.05）. 3. The similarity analysis showed that the dice similarity coefficient（Cs） between individuals in cesarean section group was higher than that in vaginal delivery group on 3^th day after birth, and the Cs lower than（or equal to）0.30 took up 21.4% of the total Cs in cesarean section group and 76.0% in vaginal delivery group,which had all significant differences between 2 groups （P 〈 0.05 ）. On 5＇h day after birth,there were no obvious differences between groups in Cs total level and its distribution. Conclusions The intra - group similarity of gut bacterium structure is higher in cesarean section group than in vaginal delivery group at 3^th day after birth, while the species and quantity in cesarean section group are fewer than those in vaginal delivery group at 5^th day after birth. Mode of delivery does affect the early stage of intestinal bacterial colonization. PCR - DGGE analysis resulted in adynamic description of intestinal bacterial coloniza- tion in the neonates and allowed visualization of bacteria that were difficult to cultivate or to detect by other methods.
出处
《实用儿科临床杂志》
CAS
CSCD
北大核心
2012年第19期1491-1494,共4页
Journal of Applied Clinical Pediatrics
关键词
新生儿
肠道菌群
分娩方式
变性梯度凝胶电泳
neonate
intestinal microbiota
mode of delivery
denaturing gradient gel electrophresis
