Farzi Nastaran, Behzad Catherine, Hasani Zahra, Alebouyeh Masoud, Zojaji Homayoun, Zali Mohammad Reza
Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Gastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Folia Microbiol (Praha). 2019 Mar;64(2):143-151. doi: 10.1007/s12223-018-0637-9. Epub 2018 Aug 10.
Mixed infections and heteroresistance of Helicobacter pylori contribute to decreased efficacy of treatments. This study aimed to investigate frequency of clarithromycin heteroresistance and its link with mixed infections, medication history, and disease severity. A total of 40 pairs of H. pylori strains were isolated from the antrum and corpus of 97 patients. Susceptibility of the strains to clarithromycin was measured by agar dilution method. Site-specific mutations of 23S rRNA at A2143G, A2142G, and A2142C positions were analyzed by PCR and genomic relatedness of pairs of the strains was determined by random amplified polymorphic DNA (RAPD)-PCR. The results showed a prevalence of 35% (14/40) clarithromycin resistance. Diversity of the antrum and corpus isolates in resistance to clarithromycin was detected among 17.5% (7/40) of the patients. Similarly, diversity in MIC value was also detected in two patients infected with the sensitive strains. Significant difference in frequency of resistance was detected among patients with peptic ulcer disease (PUD) (MIC90 32 μg/mL) and severe gastritis (MIC90 16 μg/mL), compared with those who suffered from non-ulcer dyspepsia (NUD) (MIC90 8 μg/mL) and chronic gastritis (MIC90 0.25 μg/mL). MIC values showed 8-32 folds increased levels in the corpus. A2142G, A2143G, and A2142C mutations were detected in three, two, and two patients, respectively, but not observed in 46% of the resistant strains. RAPD-PCR fingerprints showed identical molecular patterns for the isolates of the corpus and antrum in each patient. In conclusion, microevolution of H. pylori strains during chronic infection, rather than mixed infection, and inappropriate medication appear to be main reasons of treatment failure in adults.
幽门螺杆菌的混合感染和异质性耐药会导致治疗效果下降。本研究旨在调查克拉霉素异质性耐药的频率及其与混合感染、用药史和疾病严重程度的关系。从97例患者的胃窦和胃体中总共分离出40对幽门螺杆菌菌株。采用琼脂稀释法测定菌株对克拉霉素的敏感性。通过PCR分析23S rRNA在A2143G、A2142G和A2142C位点的位点特异性突变,并通过随机扩增多态性DNA(RAPD)-PCR确定菌株对之间的基因组相关性。结果显示克拉霉素耐药率为35%(14/40)。在17.5%(7/40)的患者中检测到胃窦和胃体分离株对克拉霉素耐药性的差异。同样,在两名感染敏感菌株的患者中也检测到MIC值的差异。与患有非溃疡性消化不良(NUD)(MIC90 8μg/mL)和慢性胃炎(MIC90 0.25μg/mL)的患者相比,消化性溃疡疾病(PUD)(MIC90 32μg/mL)和重度胃炎(MIC90 16μg/mL)患者的耐药频率存在显著差异。胃体中的MIC值升高了8 - 32倍。分别在3例、2例和2例患者中检测到A2142G、A2143G和A2142C突变,但在46%的耐药菌株中未观察到。RAPD-PCR指纹图谱显示每位患者胃体和胃窦分离株的分子模式相同。总之,慢性感染期间幽门螺杆菌菌株的微进化,而非混合感染和不适当用药,似乎是成人治疗失败的主要原因。