本文選題：新生兒壞死性小腸結(jié)腸炎 + 變形梯度凝膠電泳　； 參考：《重慶醫(yī)科大學(xué)》2012年碩士論文

【摘要】：目的 新生兒壞死性小腸結(jié)腸炎（NEC, neonatal necrotizingenterocolitis）是新生兒特別是早產(chǎn)兒的急腹癥，臨床以腹脹、嘔吐、便血、嚴(yán)重休克為表現(xiàn)，X線以腸壁囊樣積氣為特征。本病在國(guó)內(nèi)的病死率高，尤其是早產(chǎn)兒,嚴(yán)重影響新生兒健康。NEC是多因素的疾病，細(xì)菌感染是其中一個(gè)不可缺少的致病因素。有研究表明，NEC未被發(fā)現(xiàn)在無(wú)菌的動(dòng)物模型上,說(shuō)明了無(wú)菌無(wú)NEC。有假說(shuō)表明新生兒腸道異常細(xì)菌的定植是NEC發(fā)生的一個(gè)重要因素。但目前國(guó)內(nèi)外對(duì)NEC腸道細(xì)菌感染的細(xì)菌成分和動(dòng)態(tài)性變化并無(wú)清晰的了解。本實(shí)驗(yàn)采用PCR結(jié)合DGGE來(lái)觀察NEC腸道微生態(tài)成分和動(dòng)態(tài)變化，為NEC疾病的發(fā)病機(jī)制和臨床防治提供細(xì)菌學(xué)理論基礎(chǔ)。 材料與方法 從重慶市兒童醫(yī)院新生兒病房2009年8月份-2009年12月份共收治32例，其中NEC組和對(duì)照組各16例。NEC組根據(jù)實(shí)用兒科學(xué)診斷標(biāo)準(zhǔn)收取，每例均有x片（+）及大便隱血（+）。根據(jù)國(guó)際Walsh andKliegman的BELL對(duì)于NEC疾病的分期標(biāo)準(zhǔn),均為Ⅰ期(輕度)。對(duì)照組根據(jù)基礎(chǔ)疾病以及臨床特征：性別，胎齡，生產(chǎn)方式，喂養(yǎng)方式，出生體重，抗生素使用時(shí)間來(lái)盡量匹配。取其新鮮大便，提腸道細(xì)菌總DNA，用16s rDNA V3可變區(qū)引物進(jìn)行聚合酶鏈反應(yīng)（PCR，polymerase chain reaction）擴(kuò)增，然后變性梯度凝膠電泳（DGGE，denaturing gradient gel electrophoresis），割膠回收細(xì)菌DNA測(cè)序，進(jìn)行腸道細(xì)菌多樣性分析包括豐富度（richness，S）和Shannon index（H′）指數(shù)，序列比對(duì)分析。 結(jié)果 變形梯度凝膠電泳結(jié)果顯示：13/16（即81.25%）的NEC組跟對(duì)照組比較，腸道細(xì)菌菌落的Shannon指數(shù)相對(duì)較高(NEC組vs對(duì)照組,p0.05)，分布不均，波動(dòng)明顯，細(xì)菌的多樣性高于對(duì)照組，，細(xì)菌菌落多樣性特征存在一定的差異。NEC組的Shannon指數(shù)在發(fā)病初期明顯增多,隨著疾病的治療與恢復(fù)，呈先降后升的趨勢(shì)。DGGE膠回收測(cè)序顯示新生兒腸道優(yōu)勢(shì)菌群以肺炎克雷伯桿菌屬，大腸埃希氏菌屬，腸球菌屬為主，NEC組與對(duì)照組種類沒(méi)有明顯區(qū)別，存在數(shù)量上的差異。3/16（18.75%）趨勢(shì)不明顯，shannon index差異不大。 結(jié)論 實(shí)驗(yàn)表明，NEC患兒的腸道菌群失衡可能是NEC發(fā)病的腸道細(xì)菌感染的重要因素。
[Abstract]:Purpose Neonatal necrotizing enterocolitis (neonatal necrotizing enterocolitis) is an acute abdomen of newborns, especially premature infants. It is characterized by abdominal distension, vomiting, hematochezia and severe shock. The mortality of this disease is high in China, especially in premature infants. NEC is a multifactor disease, and bacterial infection is one of the indispensable pathogenic factors. Some studies have shown that NEC was not found in sterile animal models, indicating aseptic NECs. It is hypothesized that the colonization of abnormal intestinal bacteria in neonates is an important factor in the development of NEC. However, there is no clear understanding of the changes of bacterial composition and dynamics of NEC enterobacterial infection at home and abroad. In this study, PCR and DGGE were used to observe the microecological composition and dynamic changes of intestinal tract of NEC, which provided a theoretical basis for the pathogenesis of NEC disease and its clinical prevention and treatment. Materials and methods From August 2009 to December 2009, 32 cases were treated in the neonatal ward of Chongqing Children's Hospital. Among them, 16 cases in the NEC group and 16 cases in the control group were collected according to the diagnostic criteria of practical pediatrics. Each case had X ray and fecal occult blood. According to the international standard of BELL of Walsh andKliegman for staging of NEC disease, all of them were stage 鈪

本文編號(hào)：1835663