Clarithromycin resistance and point mutations in the 23S rRNA gene in Helicobacter pylori isolates from Malaysia
Clarithromycin resistance and point mutations in the 23S rRNA gene in Helicobacter pylori isolates from Malaysia
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DOI：
10.1111/j.1751-2980.2010.00423.x
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发表时间：
2010-04-01
影响因子：
3.5
通讯作者：
Goh, Khean-Lee
中科院分区：
文献类型：
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作者：
Ho, Sin-Loong;Tan, Eng Lai;Goh, Khean-Lee
目的：确定幽门螺杆菌（H。pylori）菌株在马来西亚胃十二指肠疾病患者中的分布情况，采用限制性片段长度多态性（RFLP）技术对23 SrRNA的V区进行分析。pylori阳性患者进行胃镜检查。DNA提取后，使用引物Hp 23 -1和Hp 23 -2进行PCR扩增，引物Hp 23 -1和Hp 23 -2位于细菌23 S rRNA肽基转移酶（Hp 23 S片段）内425 bp区域的侧翼。23 S rRNA基因突变的分析是基于两种限制性内切酶的限制性位点的产生：BbsI和BsaI，这对应于克拉霉素耐药性的碱基替换特征，从A到G的位置2142和2143，分别。仅从105个样品中PCR扩增出大小为425 bp的片段，其对应于从23 S rRNA的结构域V扩增的预期片段。扩增子随后受到BbsI和BsaI的限制。仅1例（0.95%）样本存在BbsI突变（A2142 G碱基替换），2例（1.90%）样本存在BsaI突变（A2143 G碱基替换）。结论：PCR-RFLP是一种快速、准确的检测耐药菌株的方法。幽门螺杆菌克拉霉素。使用这种方法，在我们当地的马来西亚菌株中检测到克拉霉素耐药的低流行率。这预示着克拉霉素作为一线药物继续用于治疗和根除H。幽门感染
OBJECTIVE:To determine the prevalence of primary clarithromycin resistance amongst Helicobacter pylori (H. pylori) strains in Malaysian patients with gastroduodenal diseases, by using restriction fragment length polymorphism (RFLP) in domain V of 23S rRNA.METHODS:Gastric biopsies were obtained from H. pylori positive patients undergoing gastroscopy. DNA extraction was followed by PCR amplification using the primers Hp23-1 and Hp23-2 flanking a region of 425bp within the bacterial 23S rRNA peptidyltranferase (Hp23S fragment). Analysis of the 23S rRNA gene mutations is based on the generation of restriction sites for two restriction enzymes: BbsI and BsaI, which correspond to the base substitutions characteristic of clarithromycin resistance from A to G at positions 2142 and 2143, respectively.RESULTS:Gastric biopsy samples were obtained from 107 patients. A fragment of size 425bp corresponding to that expected from amplification of domain V of 23S rRNA was PCR-amplified from only 105 samples. The amplicon was subsequently subjected to restriction by BbsI and BsaI. Only 1 sample (0.95%) had the BbsI mutation (base substitution at A2142G) and 2 samples (1.90%) the BsaI mutation (base substitution at A2143G). Thus 3 of 105 (2.9%) samples harbored clarithromycin resistant strains.CONCLUSION:In our experience, PCR-RFLP is a rapid and precise method to detect the resistance of H. pylori to clarithromycin. Using this method, a low prevalence of clarithromycin resistance was detected in our local Malaysian strains. This augurs well for the continued use of clarithromycin as a first line drug in the treatment and eradication of H. pylori infection.