PCR-RFLP detection of point mutations A2143G and A2142G in 23S rRNA gene conferring resistance to clarithromycin in Helicobacter pylori strains.

Klesiewicz, Karolina; Nowak, Paweł; Karczewska, E; Skiba, Iwona; Wojtas-Bonior, Izabela; Sito, E; Budak, Alicja

doi:10.18388/abp.2014_1901

Cited by 36 publications

(44 citation statements)

References 36 publications

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“…In our study the A2142G mutation was the commonest among the tested samples while the A2143G mutation was not found among the isolates, contrasting with other authors who reported that the commonest mutations were A2143G and A2142G found in equal percentages among the tested isolates 14 . Acosta et al .…”

Section: Discussioncontrasting

confidence: 99%

GENETIC MUTATIONS AFFECTING THE FIRST LINE ERADICATION THERAPY OF Helicobacter pylori-INFECTED EGYPTIAN PATIENTS

Ramzy

El-Garem

Hamza

et al. 2016

Rev. Inst. Med. trop. S. Paulo

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SUMMARYIntroduction: Several genetic mutations affect the first-line triple therapy for Helicobacter pylori. We aimed to study the most common genetic mutations affecting the metronidazole and clarithromycin therapy for H. pylori-infected Egyptian patients.Patients and Methods: In our study, we included 100 successive dyspeptic patients scheduled for diagnosis through upper gastroscopy at Cairo's University Hospital, Egypt. Gastric biopsies were tested for the presence of H. pylori by detection of the 16S rRNA gene. Positive biopsies were further studied for the presence of the rdxA gene deletion by Polymerase Chain Reaction (PCR), while clarithromycin resistance was investigated by the presence of nucleotide substitutions within H. pylori 23S rRNA V domain using MboII and BsaI to carry out a Restricted Fragment Length Polymorphism (RFLP) assay.Results: Among 70 H. pylori positive biopsies, the rdxA gene deletion was detected in 44/70 (62.9%) samples, while predominance of the A2142G mutations within the H. pylori 23S rRNA V domain was evidenced in 39/70 (55.7%) of the positive H. pylori cases. No statistically significant difference was found between the presence of gene mutations and different factors such as patients 'age, gender, geographic distribution, symptoms and endoscopic findings.Conclusion: Infection with mutated H. pylori strains is considerably high, a finding that imposes care in the use of the triple therapy to treat H. pylori in Egypt, since the guidelines recommend to abandon the standard triple therapy when the primary clarithromycin resistance rate is over 20%1.

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Section: Discussioncontrasting

confidence: 99%

GENETIC MUTATIONS AFFECTING THE FIRST LINE ERADICATION THERAPY OF Helicobacter pylori-INFECTED EGYPTIAN PATIENTS

Ramzy

El-Garem

Hamza

et al. 2016

Rev. Inst. Med. trop. S. Paulo

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“…These mutations prevent the macrolide from binding. Moreover, other mutations such as A2115G, G2141A, C2147G, T2190C, C2195T, A2223G and C2694A have been implicated in the development of clarithromycin resistance, although their precise role in the mechanism of resistance remains unclear …”

Section: Antibiotic Resistance Mechanismsmentioning

confidence: 99%

Review article: the global emergence ofHelicobacter pyloriantibiotic resistance

Thung

Aramin

Vavinskaya

et al. 2015

Aliment Pharmacol Ther

594

562

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SummaryBackground Helicobacter pylori is one of the most prevalent global pathogens and can lead to gastrointestinal disease including peptic ulcers, gastric marginal zone lymphoma and gastric carcinoma.AimTo review recent trends in H. pylori antibiotic resistance rates, and to discuss diagnostics and treatment paradigms.MethodsA PubMed literature search using the following keywords: Helicobacter pylori, antibiotic resistance, clarithromycin, levofloxacin, metronidazole, prevalence, susceptibility testing.ResultsThe prevalence of bacterial antibiotic resistance is regionally variable and appears to be markedly increasing with time in many countries. Concordantly, the antimicrobial eradication rate of H. pylori has been declining globally. In particular, clarithromycin resistance has been rapidly increasing in many countries over the past decade, with rates as high as approximately 30% in Japan and Italy, 50% in China and 40% in Turkey; whereas resistance rates are much lower in Sweden and Taiwan, at approximately 15%; there are limited data in the USA. Other antibiotics show similar trends, although less pronounced.ConclusionsSince the choice of empiric therapies should be predicated on accurate information regarding antibiotic resistance rates, there is a critical need for determination of current rates at a local scale, and perhaps in individual patients. Such information would not only guide selection of appropriate empiric antibiotic therapy but also inform the development of better methods to identify H. pylori antibiotic resistance at diagnosis. Patient‐specific tailoring of effective antibiotic treatment strategies may lead to reduced treatment failures and less antibiotic resistance.

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“…With the extensive implementation of eradication therapy, the resistance of H. pylori to antibiotics has been increasing year by year. 5 Other new mutation sites including T2115G, G2141A, T2190C, C2195T, A2223G, C2694A, 6 and A2144G 7 have also been mentioned in other areas. Some changes of H. pylori genes could result in the loss of antibiotics targets and thus induce bacterial resistance.…”

Section: Introductionmentioning

confidence: 97%

Molecular detection of H. pylori antibiotic‐resistant genes and molecular docking analysis

et al. 2019

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To explore the mutation characteristics of H. pylori resistance-related genes to antibiotics of clarithromycin (CAM), levofloxacin (LVX) and metronidazole, 23SrRNA, gyrA, gyrB, rdxA, and frxA genes were amplified and sequenced, respectively. Molecular docking study was performed to explore molecular interactions between chemotherapeutic agents and target proteins. In the CAM-resistant strains, the mutation rate in site A2143G was 74.2% (n = 23). The interactions in sites of G1949A, C1953T, and G2211T with CAM were weaker after mutation. In the LVX-resistant strains, the mutation rates in 87 (N to K/I) and 91 (D to N/Y/G) of gyrA were 28.6% (n = 16) and 12.5% (n = 7), respectively. We could infer by docking studies that N87 K/I, D91Y/G/N, D99N, and D143E mutations in GyrA protein all had weakened interaction with LVX. The mutation types of RdxA protein consisted of protein truncation caused by premature stop codons (n = 26, 33.3%), frameshift mutations (n = 8, 10.3%), FMN-binding sites (n = 16, 20.5%), and the others (n = 11, 14.1%).Docking analysis showed that some mutations in those four types of RdxA protein could reduce interaction with MNZ, and play a significant role in antibiotic resistance. What's more, we performed natural transformation with some of these mutated DNA fragments to see if they really impact susceptibility to antibiotics. Our study suggested that in addition to the reported mutation sites, H. pylori antibiotic resistance in this region may also be associated with changes in some new sites. Our study provides novel ideas and methods for the identification of H. pylori resistancerelated mutations, and also clues and basis for further investigation on specific molecular mechanism.

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PCR-RFLP detection of point mutations A2143G and A2142G in 23S rRNA gene conferring resistance to clarithromycin in Helicobacter pylori strains.

Cited by 36 publications

References 36 publications

GENETIC MUTATIONS AFFECTING THE FIRST LINE ERADICATION THERAPY OF Helicobacter pylori-INFECTED EGYPTIAN PATIENTS

GENETIC MUTATIONS AFFECTING THE FIRST LINE ERADICATION THERAPY OF Helicobacter pylori-INFECTED EGYPTIAN PATIENTS

Review article: the global emergence ofHelicobacter pyloriantibiotic resistance

Molecular detection of H. pylori antibiotic‐resistant genes and molecular docking analysis

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