粪便基因型检测幽门螺杆菌对克拉霉素耐药性的研究  

Detection of Helicobacter pylori Resistance to Clarithromycin by Fecal Molecular Genotyping

作　　者：秦幼娟[1] 周晓颖[1] 赵冰[1] 刘伟[1] 潘晓林[1] 张国新[1] 

机构地区：[1]南京医科大学第一附属医院消化科,210029

出　　处：《胃肠病学》2013年第12期747-749,共3页Chinese Journal of Gastroenterology

基　　金：国家自然科学基金(No.81270476;81072032)资助

摘　　要：幽门螺杆菌(Hp)耐药情况日趋严重,选择快速、敏感、价廉的分子生物学技术对Hp耐药进行检测具有重要的临床意义。目的:评价检测粪便Hp基因突变对诊断克拉霉素耐药的有效性,并探讨cagA基因与耐药的相关性。方法:纳入74例13C-尿素呼气试验阳性患者,采集其新鲜粪便标本,提取粪便DNA,采用巢式PCR法扩增Hp 23S rRNA,采用PCR-RFLP法检测限制性内切酶BbsⅠ、BceAⅠ、BsaⅠ对23S rRNA扩增产物的酶切情况,采用PCR法扩增cagA基因。结果:74例患者的粪便标本中,60例扩增出Hp 23S rRNA 367 bp片段,其中17例可被BsaⅠ酶切,60例均未被BbsⅠ、BceAⅠ酶切。cagA阳性、阴性表达者的23S rRNA突变率相比差异无统计学意义(P>0.05)。结论:通过粪便基因型检测Hp对克拉霉素耐药是快速、简便的方法。江苏地区Hp对克拉霉素的耐药机制主要为23S rRNA A2143G突变。cagA基因与Hp对克拉霉素耐药不相关。Background： The rate of Helicobacter pylori （Hp） resistance to antibiotics is increasing, it is important to choose a fast, sensitive and inexpensive molecular biology technology to test the resistance of Hp. Aims： To evaluate the efficiency of fecal molecular genotyping to detect Hp resistance to clarithromycin and investigate the relationship between cagA and clafithromycin resistance. Methods： A total of 74 Hp-infected patients diagnosed by ~3 C-urea breath test were enrolled. Stool samples of the patients were collected and DNA was extracted. Hp 23S rRNA was amplified by nested PCR. The fragments of Hp 23S rRNA digested by restriction enzyme Bbs I , BceA I and Bsa I were detected by PCR-RFLP. cagA gene was amplified by PCR. Results： 367 bp fragment of Hp 23S rRNA could be amplified in 60 stool samples, in which 17 could be digested by Bsa I , none could be digested by Bbs I or BceA I. No significant difference of mutation rate of 23S rRNA was seen between cagA-positive and cagA-negative cases （P 〉0.05 ）. Conclusions： Detection of Hp resistance to clarithromycin by fecal molecular genotyping is a fast and convenient method. Hp resistance to clarithromycin is due to A2143G mutation in 23S rRNA in Jiangsu area. There is no relationship between cagA gene and clarithromycin resistance.

关 键 词：幽门螺杆菌 克拉霉素 耐药性 粪便 基因突变 

分 类 号：R57[医药卫生—消化系统]