摘要
目的:通过对幽门螺杆菌(Helicobacter pylori,HP)23s rRNA基因突变位点分析,观察23s rRNA各突变点与克拉霉素耐药的关系。方法:分离培养HP、提取DNA、扩增其23s rRNA基因片段及测序;运用Clustal W软件进行序列比对找出突变点。选用GenBank中已全基因组测序的35株HP,根据其mutY、ppa、efp和ureI 4个管家基因建立Neighbour-joining树,根据进化树结果将35株HP分为欧非和东亚菌群;观察各突变点与菌群分布的关系。结果:与耐药有关的A2143G突变率为32.8%,T2182C突变率为90.6%但与耐药无关,其碱基分布具有菌群差异:东亚菌群以C碱基为主,欧非菌群以T为主(P<0.05),其他可能与耐药有关的各位点分别为A2223G(6.25%)、T2190C(1.6%)、C2195T(1.6%)。结论:青岛地区HP克拉霉素耐药率为32.8%并表现为23s rRNA基因A2143G突变,而2182位点突变是由于菌群分布差异导致的与克拉霉素耐药无关。
Objective： To investigate the mutation sites of the 23s rRNA gene of Helicobacter pylori（HP）,and observe the relationship between the mutation sites and HP clarithromycin resistance.Methods： HP strains were isolated from gastric mucosa biopsies,then DNA was extracted,and 23s rRNA gene was amplifed by PCR and sequenced.The gene sequences were aligned by using the Clustal W software in order to find the mutation sites,and the Neighbour-joining tree were built by using four housekeeping genes（mutY,ppa,efp and ureI） of 35 HP strains which from the GenBank.The 35 HP strains were divided into EastAsia group and EuroAfrica group according to the Neighbour-joining tree and the distribution characteristic of mutation sites between the different groups was detected.Results： The prevalence of A2143G was 32.8 % which represent the clarithromycin resistance.T2182C（90.6 %） mutation was not related with the clarithromycin resistance,but with the group difference,in the EastAsia group the 2182 site is mainly expressed as C,while in the EuroAfrica group was T（P0.05）.Other mutations such as A2223G（6.25 %）,T2190C（1.6 %）,and C2195T（1.6 %） were also detected.Conclusion： The prevalence of HP clarithromycin resistance is 32.8 % mainly expressed as A2143G mutation,the T2182C mutation is not related with the HP clarithromycin resistance but with the difference of HP distribution in the world.
出处
《现代生物医学进展》
CAS
2013年第13期2468-2471,共4页
Progress in Modern Biomedicine
