2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649037摘要：成人的腸道中含有各種共生菌，每公克大便約有1014 細菌。胎兒在子宮內每天吞進的是無菌羊水，因此它的腸道是無菌的狀態；而剛出生嬰兒的腸道是如何轉變成像大人一樣的型態？嬰幼兒腸道菌種、數量的演進與很多因素有關。腸道是人體最大的免疫器官。它接受無數的細菌從口腔中隨著食物進入，有些細菌會被排出、有些則被腸道細胞所認識而產生耐受性。過敏性疾病的發生率有所謂的「衛生理論」，因此我們相信腸內菌的種類與過敏性疾病有關。傳統研究腸內菌是靠培養法，其有一定的限制與難度、也無法定量分析。較新的方法則是針對細菌共有的16s-RNA/DNA 以PCR 將其偵測並定序定量（PCR-DGGE）。藉以提升腸內菌偵測、鑑別與定量的準確性。本研究是採前瞻、長期性收集從出生嬰兒的胎便，健兒門診（1、2、6、12、18、24、36 個月）嬰兒的糞便直到三歲；臨床上紀錄其各種健康狀態與餵食種類。如果追蹤期間嬰幼兒發生疾病時，會再收集糞便。以PCR-DGGE 方法檢測糞便中細菌種類與數量。在三歲時並檢測其各種過敏性指標。藉此可以知道國內嬰幼兒腸內菌的建立與各種情境之關係；並藉以了解嬰幼兒發生疾病時，其腸內菌在疾病前、中、後之型態改變。最後，本研究也想探討嬰幼兒腸內菌的演化與三歲時，過敏性疾病其相關性。<br> Abstract: There are large amounts of commensal microbiota in adult gastrointestinal (GI)tract, which harbor about 1014 bacteria per gram of fecal mass. When in uteri, fetusswallows sterile amniotic fluid. Hence, the GI tract of fetus is supposed to be sterilealso. It is interesting to know how the newly born infant acquits his/her first microflorain the gut and how the evolution of microbiota residence in newborn’s intestinechanges to adult type colonization. The origin of the bacteria colonizing the neonatalGI tract is supposed to be affected by mode of delivery, mode of feeding, differenthospitalization settings or maternal health condition. The different status of hygienewill also influence the acquisition of the microbiota to inhabit the infant’s intestine.Previous reports have documented the different microbiota communities betweendeveloping and developed countries.Besides, the GI tract is the largest immunological organ in human body. Largeamount of bacteria are swallowed through the mouth accompany with food ingestedevery day. Some bacteria are expelled; some residence and play a role in facilitatingoral tolerance. The intricacy of the immune modulation between tolerance andallergy is complex and not fully understood. We can hypothesize that the microbiotain the developing gut are closely related to the development of allergy disease.Traditionally, studies for the intestinal flora were based on the stool culture(culture-based). The sensitivity of culture-based study had large variability dependingon the cultural medium and environment. With the advanced technology, DNA-basestudy stands the major role in exploring the intestinal microbiota compositions. Priorstudies of bacterial evolution and phylogenetics have paved the foundation forsequencing-based 16S rRNA (or16S rDNA) to identify microbes. With the ability toamplify 16S rDNA from a whole bacteria community, such a technique is multiplexedto a new horizon ready for more detailed and comprehensive analyses. Denaturinggradient gel electrophoresis (DGGE) has been used as a tool for profiling complexmicrobial populations without the bias of cultural analysisThis prospective and longitudinal study is designed to follow and to collect thestool specimen from birth to 3 years old at the time of 1, 2, 6, 12, 18, 24 and 36 m/o. Ifthe enrolled infant gets common infection such as gastroenterocolitis, acutebronchiolitis or necrotizing enterocolitis, we’ll collect another stool during the course ofdisease and after the recovery phase. The stool will be processing with PCR-DGGEmethod to detect all microbiota in stool. We’ll also check the immune status of theenrolled infant at 3 years old.Through this study, we want to get the information about the acquisition of microbiotain the intestine of our infants, the evolution of the microbiota from infant to children.We can also understand the relationships between the intestinal microbiota andgastroenterocolitis, acute bronchiolitis or necrotizing enterocolitis and the allergicdiseases.腸道共生菌時間溫度梯度凝膠電泳commensal microbiotatemporal temperature gradient gel electrophoresis