埃及开罗幽门螺杆菌23S rRNA基因V结构域的突变
Mutations affecting domain V of the 23S rRNA gene in Helicobacter pylori from Cairo, Egypt.
作者信息
Ghaith Doaa, Elzahry Mohammad, Mostafa Gehan, Mostafa Sally, Elsherif Rasha, Ramzy Iman
机构信息
a Clinical and Chemical Pathology Department, Faculty of Medicine , Cairo University , Egypt.
b Endemic Medicine and Hepatology Department, Faculty of Medicine , Cairo University , Egypt.
出版信息
J Chemother. 2016 Oct;28(5):367-70. doi: 10.1179/1973947815Y.0000000067. Epub 2016 Jun 28.
BACKGROUND
Clarithromycin is a main component of the recommended first-line triple therapy for Helicobacter pylori in Egypt. We aimed in our study to investigate the prevalence of clarithromycin-resistant H. pylori strains due to the point mutations at domain V of the H. pylori 23S rRNA among the Egyptian population using the polymerase chain reaction/restricted fragment length polymorphism (PCR/RFLP) assay.
METHODS
Gastric biopsies obtained from 100 dyspeptic patients who consecutively attended at Cairo University Hospital during the period from January to November 2013 were subjected to PCR/RFLP in order to detect the point mutations at domain V of the H. pylori 23S rRNA associated with clarithromycin resistance. The PCR amplicon of the 23S H. pylori rRNA is restricted with MboII for detection of A2142G mutation and with BsaI for A2143G mutation.
RESULTS
The prevalence of H. pylori infection among 100 patients was 70%; clarithromycin resistance was detected in 39/70 (57.7%) of positive H. pylori isolates. Occurrence of 23S rRNA A2142G mutations resulted in two DNA fragments (418 and 350 bp) by PCR-RFLP; on the other hand, no A2143G mutations were detected.
CONCLUSIONS
The high prevalence of clarithromycin resistance (57.7%) caused by A2142G mutations at domain V of the H. pylori 23S rRNA may mandate changing of the standard clarithromycin-containing triple therapy. The PCR/RFLP assay was a rapid and accurate method for molecular detection of H. pylori infection in addition to determination of different nucleotide mutations causing clarithromycin resistance.
背景
克拉霉素是埃及推荐用于幽门螺杆菌一线三联疗法的主要成分。我们在本研究中的目的是,使用聚合酶链反应/限制性片段长度多态性(PCR/RFLP)分析法,调查埃及人群中因幽门螺杆菌23S rRNA结构域V点突变导致的克拉霉素耐药幽门螺杆菌菌株的流行情况。
方法
对2013年1月至11月期间在开罗大学医院连续就诊的100例消化不良患者获取的胃活检组织进行PCR/RFLP分析,以检测与克拉霉素耐药相关的幽门螺杆菌23S rRNA结构域V的点突变。幽门螺杆菌23S rRNA的PCR扩增产物用MboII酶切以检测A2142G突变,用BsaI酶切以检测A2143G突变。
结果
100例患者中幽门螺杆菌感染率为70%;在70例幽门螺杆菌阳性分离株中,检测到39株(57.7%)对克拉霉素耐药。通过PCR-RFLP分析,23S rRNA A2142G突变导致产生两个DNA片段(418和350 bp);另一方面,未检测到A2143G突变。
结论
幽门螺杆菌23S rRNA结构域V的A2142G突变导致克拉霉素耐药的高流行率(57.7%)可能需要改变含克拉霉素的标准三联疗法。PCR/RFLP分析法是一种快速准确的分子检测幽门螺杆菌感染的方法,此外还可用于确定导致克拉霉素耐药的不同核苷酸突变。