项目名称： 肠杆菌科细菌blaNDM-1基因跨种属转移及耐药进化研究

项目编号： No.81471986

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 杜小幸

作者单位： 浙江大学

项目金额： 80万元

中文摘要： 我国首例blaNDM-1阳性肠杆菌科细菌报道于2011年，短短3年间该菌在河南、云南、湖南、山东及浙江等地出现，有很快广泛流行的趋势。急需明确blaNDM-1基因如何进入肠杆菌科细菌，后者又如何发展成为威胁严重的超级细菌。本课题组前期已收集35株携带blaNDM-1基因的肠杆菌科细菌，其耐药基因均位于质粒上。本研究拟通过接合实验，明确携带blaNDM-1质粒在细菌种内及种间的转移能力；通过质粒的高通量测序及序列分析，明确携带blaNDM-1质粒的序列特点；通过转化子连续传代过的适应性代价研究，探寻耐药菌发展的规律；运用基因组DNA高通量测序、基因敲除及回补实验，验证介导传代转化子适应性回复的补偿突变及其功能。阐明blaNDM-1基因在肠杆菌科细菌种内和不同种属间快速扩散的机制及携带blaNDM-1基因肠杆菌科细菌耐药进化的规律，为控制blaNDM-1基因在不同种属间的传播提供科学依据。

中文关键词： blaNDM-1；肠杆菌科细菌；水平基因转移；耐药进化；适应性代价

英文摘要： blaNDM-1-positive Enterobacteriaceae isolate was firstly reported in 2009 in China, while it rapidly spread multiple provinces and cities within recent years, such as Henan, Yunnan, Hunan, Shandong and Zhejiang province, which indicates it would be a epidemic threaten. It is such an important issue to figure out how Enterobacteriaceae isolates obtain the blaNDM-1 gene and develope to such pathogens, known as Superbug, threatening to public health. In this study, 35 blaNDM-1-positive Enterobacteriaceae isolates are collected and have confirmed all of which are located on plasmids. We are going to evaluate the transfer ability of blaNDM-1 plasmid intra-species by using filt-mating experiment, investigate the sequence characteristics of the blaNDM-1 plasmid using high-throughput sequencing technology followed by sequence alignment. To reveal evolution regularity of the transconjugants during serial passage, fitness-cost values and compensatory mutations of chromosome will then be detected by fitness-cost experiment and whole genomic sequencing, respectively. The function of those chromosome mutations will be testified by gene knockout and genetic complemantation experiment. In this study, we hope to clarify the dissemination mechanism of the blaNDM-1 gene intra-species, reveal the evolution regularity of blaNDM-1-positive Enterobacteriaceae and finally find effective provention and control measures to this clinical pathogen.

英文关键词： blaNDM-1;Enterobacteriaceae;horizontal gene transfer;molecular epidemiology;fitness cost