吉林大学学报(医学版) ›› 2022, Vol. 48 ›› Issue (3): 630-637.doi: 10.13481/j.1671-587X.20220311

• 基础研究 • 上一篇    

幽门螺杆菌对克拉霉素、甲硝唑和左氧氟沙星的耐药率及其相关耐药基因突变特征

宋顺佳1,王鑫莹1,姜菲菲1,贾慧建1,赵远1,杨志平2,孙丽媛1(),赵云冬1()   

  1. 1.北华大学医学技术学院分子生物教研室,吉林 吉林 132013
    2.北华大学附属医院消化内科,吉林 吉林 132011
  • 收稿日期:2021-09-23 出版日期:2022-05-28 发布日期:2022-06-21
  • 通讯作者: 孙丽媛,赵云冬 E-mail:jlsunliyuan@163.com;18604498530@163.com
  • 作者简介:宋顺佳(1995-),女,吉林省磐石市人,在读硕士研究生,主要从事微生物和食品及中药DNA指纹技术应用方面的研究。
  • 基金资助:
    吉林省教育厅“十三五”科学技术项目(JJKH20190657KJ);北华大学研究生创新计划项目(北华研创合字[2019]026)

Resistance rates of Helicobacter pylori to clarithromycin, metronidazole, and levofloxacin and mutation characteristics of their related drug resistance genes

Shunjia SONG1,Xinying WANG1,Feifei JIANG1,Huijian JIA1,Yuan ZHAO1,Zhiping YANG2,Liyuan SUN1(),Yundong ZHAO1()   

  1. 1.Department of Molecular Biology,School of Medical Technology,Beihua University,Jilin 132013,China
    2.Department of Gastroenterology,Affiliated Hospital,Beihua University,Jilin 132011,China
  • Received:2021-09-23 Online:2022-05-28 Published:2022-06-21
  • Contact: Liyuan SUN,Yundong ZHAO E-mail:jlsunliyuan@163.com;18604498530@163.com

摘要: 目的

检测幽门螺杆菌(Hp)对3种常见抗生素的耐药率,探讨其相关耐药基因突变形式,为阐明其耐药分子机制提供依据。

方法

选择35例确诊为慢性胃炎或消化性溃疡患者的胃黏膜标本,分离培养鉴定Hp。采用琼脂稀释法检测Hp对克拉霉素、甲硝唑和左氧氟沙星的最小抑菌浓度(MIC),根据药敏试验结果分为敏感组和耐药组。通过PCR扩增克拉霉素耐药基因23S rRNA、甲硝唑耐药基因rdxA和左氧氟沙星耐药基因gyrA,PCR纯化产物测序后分析核苷酸和氨基酸突变位点。

结果

Hp菌株培养3~5 d后,平板上可见光滑、半透明和针尖样小菌落。35株Hp均对甲硝唑和左氧氟沙星耐药,克拉霉素耐药率为80%(28/35)。与敏感菌株比较,克拉霉素耐药菌株均发生基因突变,23S rRNA基因最常见的突变位点为A2143G,其次为突变位点A2223G和A2142G+A2164G;甲硝唑耐药菌株rdxA基因主要发生错义突变,其次为移码突变和无义突变;左氧氟沙星耐药菌株gyrA基因主要发生N87K突变,其次为D91N突变、N87I、D91G及N87K+D143E突变,17.1%的耐药菌株未发生任何位点突变。

结论

Hp菌株对克拉霉素、甲硝唑和左氧氟沙星的耐药率较高,相关耐药基因突变是导致抗生素耐药的主要原因,筛选这些突变可能有助于防止抗生素耐药性及选择合适的抗菌药物根除Hp。

关键词: 幽门螺杆菌, 耐药性, 克拉霉素, 甲硝唑, 左氧氟沙星

Abstract:

Objective: To detect the drug resistance rates of Helicobacter pylori (Hp) to three common antibiotics,and to explore the mutation forms of drug resistance genes in order to provide the basis for elucidating the molecular mechanism of drug resistance.

Methods

The gastric mucosa samples from 35 patients with chronic gastritis or peptic ulcer were isolated and cultured to identify Hp.The minimal inhibitory concentrations (MIC) of Hp to clarithromycin, metronidazole and levofloxacin were detected by the agar dilution method and Hp was divided into sensitive group and resistant group according to the results of the drug sensitivity test.The clarithromycin resistance gene 23S rRNA, metronidazole resistance gene rdxA, and levofloxacin resistance gene gyrA were amplified by PCR method. The nucleotide and amino acid mutations were analyzed after the sequencing of PCR purified product.

Results

After cultured for 3-5 d, smooth, translucent, needle-like colonies could be seen on the plate of Hp strain. All 35 Hp strains were resistant to metronidazole and levofloxacin, the resistance rate to clarithromycin was the highest(80%). Compared with the sensitive strains, all the clarithromycin- resistant strains had gene mutations. The most common mutation site of the 23S rRNA gene was A2143G, followed by mutation sites A2223G and A2142G+A2164G. The rdxA gene of metronidazole resistant strains mainly had missense mutations, followed by frameshift mutations and nonsense mutations. The N87K mutation in the gyrA gene of levofloxacin resistant strains was the highest, followed by the D91N mutation, N87I, D91G, and N87K+D143E mutations,but 17.1% of the resistant strains did not have any site mutations.

Conclusion

The resistance rates of Hp to clarithromycin, metronidazole, and levofloxacin are high, and the mutations of related drug resistance genes are the main cause of the antibiotic resistance. Screening these mutations may help to prevent the antibiotic resistance and select the appropriate antimicrobials to eradicate Hp.

Key words: Helicobacter pylori, Drug resistance, Clarithromycin, Metronidazole, Levofloxacin

中图分类号: 

  • R446.5