Helicobacter pylori Eradication According to Sequencing-Based 23S Ribosomal RNA Point Mutation Associated with Clarithromycin Resistance

Background/Aims: Clarithromycin resistance in Helicobacter pylori is associated with point mutations in the 23S ribosomal RNA (rRNA) gene. We investigated the point mutations in the 23S rRNA genes of patients with clarithromycin-resistant H. pylori and compared the H. pylori eradication rates based on the point mutations. Methods: A total of 431 adult patients with H. pylori infection were recruited in Kangdong Sacred Heart Hospital in 2017 and 2018. Patients who did not have point mutations related to clarithromycin resistance and/or had clinically insignificant point mutations were treated with PAC (proton pump inhibitor, amoxicillin, clarithromycin) for seven days, while patients with clinically significant point mutations were treated with PAM (proton pump inhibitor, amoxicillin, metronidazole) for seven days. H. pylori eradication rates were compared. Results: Sequencing-based detection of point mutations identified four mutations that were considered clinically significant (A2142G, A2142C, A2143G, A2143C). The clarithromycin resistance rate was 21.3% in the overall group of patients. A2143G was the most clinically significant point mutation (84/431, 19.5%), while T2182C was the most clinically insignificant point mutation (283/431, 65.7%). The overall H. pylori eradication rate was 83.7%, and the seven-day PAM-treated clarithromycin-resistance group showed a significantly lower eradication rate than the seven-day PAC-treated nonresistance group (ITT; 55.4% (51/92) vs. 74.3% (252/339), p = 0.001, PP; 66.2% (51/77) vs. 88.4% (252/285), p = 0.0001). Conclusions: There were significantly lower eradication rates in the patients with clarithromycin-resistant H. pylori when treated with PAM for seven days. A future study comparing treatment regimens in clarithromycin-resistant H. pylori-infected patients may be necessary.

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Helicobacter pylori eradication according to sequencing-based 23S ribosomal RNA point mutation associated with clarithromycin resistance

10.21203/rs.2.12783/v1 ◽

2019 ◽

Author(s):

Seung In Seo ◽

Byoung Joo ◽

Jin Gu Kang ◽

Hyoung Su Kim ◽

Myoung Kuk Jang ◽

...

Keyword(s):

Point Mutation ◽

Ribosomal Rna ◽

Point Mutations ◽

23S Rrna ◽

Rrna Gene ◽

Clarithromycin Resistance ◽

Clinically Significant ◽

H Pylori ◽

23S Ribosomal Rna ◽

Significant Point

Abstract Background Clarithromycin resistance in Helicobacter pylori (H. pylori) is associated with point mutations in the 23S ribosomal RNA (rRNA) gene. A sequencing-based method can detect more point mutations than a polymerase chain reaction (PCR)-based method. We investigated the point mutations in the 23S rRNA genes of patients infected with clarithromycin-resistant H. pylori and compared the H. pylori eradication rates based on the identified clinically significant point mutations. Methods A total of 431 adult patients with H. pylori infection were retrospectively recruited in Kangdong Sacred Heart Hospital in 2017 and 2018. Patients who did not have point mutations related to clarithromycin resistance and/or had clinically insignificant point mutations were treated with PAC (proton pump inhibitor, amoxicillin, clarithromycin) for 7 days, while patients with clinically significant point mutations were treated with PAM (proton pump inhibitor, amoxicillin, metronidazole) for 7 days. H. pylori eradication rates were compared between the two groups. Results Sequencing-based detection of point mutations identified four mutations that were considered clinically significant (A2142G, A2142C, A2143G, A2143C), while all the other mutations were considered clinically insignificant. The clarithromycin resistance rate was 21.3% in the overall group of patients. A2143G was the most clinically significant point mutation (84/431, 19.5%), while T2182C was the most clinically insignificant point mutation (283/431, 65.7%). The H. pylori eradication rate in the overall group of patients was 83.7%, and the 7-day PAM-treated clarithromycin-resistance group showed a significantly lower eradication rate than the 7-day PAC-treated nonresistance group [ITT; 55.4% (51/92) vs. 74.3% (252/339), P=0.001, PP; 66.2% (51/77) vs. 88.4% (252/285), P=0.0001]. Conclusions There were significantly lower eradication rates in the patients with clarithromycin-resistant H. pylori as identified by the sequencing of point mutations in the 23S rRNA gene when treated with PAM for 7 days. A future study comparing treatment regimens in clarithromycin-resistant H. pylori-infected patients may be necessary.

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Eradication of Helicobacter pylori According to 23S Ribosomal RNA Point Mutations Associated With Clarithromycin Resistance

The Journal of Infectious Diseases ◽

10.1093/infdis/jit287 ◽

2013 ◽

Vol 208 (7) ◽

pp. 1123-1130 ◽

Cited By ~ 48

Author(s):

H. J. Lee ◽

J. I. Kim ◽

D. Y. Cheung ◽

T. H. Kim ◽

E. J. Jun ◽

...

Keyword(s):

Helicobacter Pylori ◽

Ribosomal Rna ◽

Point Mutations ◽

Clarithromycin Resistance ◽

23S Ribosomal Rna

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Different dose of new generation proton pump inhibitors for the treatment of Helicobacter pylori infection: A meta-analysis

International Journal of Immunopathology and Pharmacology ◽

10.1177/20587384211030397 ◽

2021 ◽

Vol 35 ◽

pp. 205873842110303

Author(s):

Wenwen Gao ◽

Xiang Zhang ◽

Yanhui Yin ◽

Shuwen Yu ◽

Lu Wang

Keyword(s):

Helicobacter Pylori ◽

Proton Pump Inhibitors ◽

Proton Pump ◽

Eradication Rate ◽

Standard Dose ◽

Statistical Significance ◽

High Dose ◽

Clarithromycin Resistance ◽

H Pylori ◽

New Generation

The evidence on whether high-dose new generation proton pump inhibitors (PPIs) including rabeprazole and esomeprazole achieve a higher eradication rate of Helicobacter pylori has not been assessed. The primary comparison was eradication and adverse events (AEs) rate of standard (esomeprazole 20 mg bid, rabeprazole 10 mg bid) versus high-dose (esomeprazole 40 mg bid, rabeprazole 20 mg bid) PPIs. Sub-analyses were performed to evaluate the eradication rate between Asians and Caucasians, clarithromycin-resistance (CAM-R) strains, and clarithromycin-sensitivity (CAM-S) strains of different dose PPIs. We conducted a literature search for randomized controlled trials comparing high-with standard-dose esomeprazole and rabeprazole for H. pylori eradication and AEs. A total of 12 trials with 2237 patients were included. The eradication rate of high-dose PPIs was not significantly superior to standard-dose PPIs regimens: 85.3% versus 84.2%, OR 1.09 (0.86–1.37), P = 0.47. The high dose induced more AEs than those of the standard dose, but didn’t reach statistical significance (OR 1.25, 95% CI: 0.99–1.56, P = 0.06). Subgroup analysis showed that the difference in eradication rate of PPIs between high- and standard-dose groups were not statistically significant both in Asians (OR 0.99, 95% CI 0.75–1.32, P = 0.97) and Caucasians (OR 1.27, 95% CI 0.84–1.92, P = 0.26). Furthermore, there were similar eradication rates in CAM-S (OR 1.2; 95% CI 0.58–2.5; P = 0.63) and CAM-R strains (OR 1.08; 95% CI 0.45–2.56; P = 0.87) between the standard-and high-dose groups. High and standard dosages of new generation of the PPIs showed similar H. pylori eradication rates and AEs as well as between Asian versus Caucasian populations, with or without clarithromycin-resistance. However, further studies are needed to confirm.

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Antimicrobial Susceptibility and Mutations Involved in Clarithromycin Resistance in Helicobacter pylori Isolates from Patients in the Western Central Region of Colombia

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00145-09 ◽

2009 ◽

Vol 53 (9) ◽

pp. 4022-4024 ◽

Cited By ~ 22

Author(s):

Adalucy Álvarez ◽

José Ignacio Moncayo ◽

Jorge Javier Santacruz ◽

Mario Santacoloma ◽

Luisa Fernanda Corredor ◽

...

Keyword(s):

Helicobacter Pylori ◽

Point Mutation ◽

Central Region ◽

Antimicrobial Susceptibility ◽

Empirical Treatment ◽

Clarithromycin Resistance ◽

H Pylori

ABSTRACT Resistance to metronidazole, clarithromycin, and amoxicillin (amoxicilline) was found in 82, 3.8, and 1.9% of 106 Helicobacter pylori isolates, respectively. No tetracycline-resistant isolates were found. In all of the clarithromycin-resistant isolates, only one point mutation was present, either A2143G or A2142G. Our results indicate that metronidazole should not be included in the empirical treatment of H. pylori infection in this region.

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Rapid screening of clarithromycin resistance in Helicobacter pylori by pyrosequencing

Journal of Medical Microbiology ◽

10.1099/jmm.0.47371-0 ◽

2007 ◽

Vol 56 (10) ◽

pp. 1370-1376 ◽

Cited By ~ 11

Author(s):

Karen-Anja Moder ◽

Franziska Layer ◽

Wolfgang König ◽

Brigitte König

Keyword(s):

Helicobacter Pylori ◽

Point Mutations ◽

Rapid Screening ◽

23S Rrna ◽

Resistance Testing ◽

Resistance Mutations ◽

Additional Time ◽

Clarithromycin Resistance ◽

H Pylori ◽

Domain V

Helicobacter pylori infections can be effectively treated with clarithromycin, a macrolide, in combination with other antibiotics, such as amoxicillin, tetracycline or metronidazole. The failure of H. pylori eradication is mainly associated with macrolide-resistant strains. Three point mutations (A2142G/C, A2143G, T2182C) in the peptidyltransferase region of domain V of the 23S rRNA have been described as being associated with clarithromycin resistance. Therefore, the determination of clarithromycin resistance by pyrosequencing was evaluated. H. pylori from 81 gastric biopsies was cultured and clarithromycin resistance was determined by Etest, as well as by pyrosequencing technology (PSQ 96 system; Biotage). The respective mutations were set in relation to the MIC measured in μg ml−1 by Etest. In this study, point mutations in positions 2142 and 2143 were associated with clarithromycin resistance. Mutations in position 2182 did not contribute to clarithromycin resistance. In addition, from 22 out of the 81 biopsies, clarithromycin resistance was determined directly without culturing H. pylori to save additional time. Identical results were obtained as compared to resistance testing with pure H. pylori strains. All results obtained by pyrosequencing were evaluated by Sanger sequencing. The data show that pyrosequencing to detect point mutation is a fast and reliable method for determining clarithromycin resistance in H. pylori, and provides the same results as the Etest.

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Antimicrobial Susceptibility of Canadian Isolates ofHelicobacter pyloriin Northeastern Ontario

Canadian Journal of Infectious Diseases and Medical Microbiology ◽

10.1155/2015/853287 ◽

2015 ◽

Vol 26 (3) ◽

pp. 137-144 ◽

Cited By ~ 8

Author(s):

Nelson F Eng ◽

Gustavo Ybazeta ◽

Katrina Chapman ◽

Nya L Fraleigh ◽

Rebecca Letto ◽

...

Keyword(s):

Antibiotic Resistance ◽

Antimicrobial Resistance ◽

Ribosomal Rna ◽

Antimicrobial Susceptibility ◽

Point Mutations ◽

Test Methods ◽

Surveillance Programs ◽

Resistant Strains ◽

H Pylori ◽

23S Ribosomal Rna

BACKGROUND:Helicobacter pyloriplays a significant role in gastritis and ulcers. It is a carcinogen as defined by the WHO, and infection can result in adenocarcinomas and mucosa-associated lymphoid tissue lymphomas. In Canada, rates of antimicrobial resistance are relatively unknown, with very few studies conducted in the past 15 years.OBJECTIVE: To examine rates of resistance in Sudbury, Ontario, compare antimicrobial susceptibility methods and attempt to determine the molecular basis of antibiotic resistance.METHODS: Patients attending scheduled visits at Health Sciences North (Sudbury, Ontario) provided gastric biopsy samples on a volunteer basis. In total, 20H pyloriisolates were collected, and antimicrobial susceptibility testing (on amoxicillin, tetracycline, metronidazole, ciprofloxacin, levofloxacin and clarithromycin) was conducted using disk diffusion and E-test methods. Subsequently, genomic DNA from these isolates was sequenced to detect mutations associated with antimicrobial resistance.RESULTS: Sixty-five percent of the isolates were found to be resistant to at least one of the listed antibiotics according to E-test. Three isolates were found to be resistant to ≥3 of the above-mentioned antibiotics. Notably, 25% of the isolates were found to be resistant to both metronidazole and clarithromycin, two antibiotics that are normally prescribed as part of first-line regimens in the treatment ofH pyloriinfections in Canada and most of the world. Among the resistant strains, the sequences of 23S ribosomal RNA andgyrA, which are linked to clarithromycin and ciprofloxacin/levofloxacin resistance, respectively, revealed the presence of known point mutations associated with antimicrobial resistance.CONCLUSIONS: In general, resistance to metronidazole, ciprofloxacin/levofloxacin and clarithromycin has increased since the studies in the early 2000s. These results suggest that surveillance programs ofH pyloriantibiotic resistance may need to be revisited or improved to prevent antimicrobial therapy failure.

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Eradication Rates in Italian Subjects Heterogeneously Managed for Helicobacter pylori Infection. Time to Abandon Empiric Treatments in Southern Europe

Journal of Gastrointestinal and Liver Diseases ◽

10.15403/jgld.2014.1121.262.itl ◽

2017 ◽

Vol 26 (2) ◽

pp. 129-137 ◽

Cited By ~ 7

Author(s):

Agostino Di Ciaula ◽

Giuseppe Scaccianoce ◽

Marino Venerito ◽

Angelo Zullo ◽

Leonilde Bonfrate ◽

...

Keyword(s):

Helicobacter Pylori ◽

Breath Test ◽

Eradication Rate ◽

Real Life ◽

Sequential Therapy ◽

Urea Breath Test ◽

Southern Europe ◽

Clarithromycin Resistance ◽

Asymptomatic Patients ◽

H Pylori

Background & Aims: H. pylori eradication is strongly affected by various factors, including the ongoing antibiotic resistance. We describe a “real life” scenario in patients managed for H. pylori-related conditions, living in a southern Italian region (Apulia), an area with clarithromycin resistance >15%.Methods: 2,224 subjects were studied in two tertiary referral centers in Apulia. Analyses included: reason for referral, H. pylori infection rates (13C-urea breath test – UBT or upper endoscopy), and eradication rates following distinct regimens previously prescribed or prospectively prescribed (such as the bismuth-based quadruple therapy Pylera®, recently marketed in Italy).Results. Over 80% of the patients were referred by family physicians (60% naïve subjects). The overall infection rate was 32.5% and it was similar in asymptomatic patients (31.1%) or with H. pylori-related symptoms/clinical conditions (34.3%). In the 987 H. pylori+ve patients receiving therapy, the overall eradication rate was 80.2% (ITT). Observed eradication rate varied greatly across different regimens: 57.1% (2nd line levofloxacin), 59.6% (unconventional), 70.7% (7-day triple), 73.2% (7-day undefined), 89% (10-day sequential) and 96.9% (ITT, 10 day Pylera®, 1st to 5th line regimens given to 227 patients).Conclusions. A heterogeneous “real life” scenario in Southern Europe shows that H. pylori+ve patients are put at risk of poor outcomes and points to the need of a susceptibility-based therapy according to guidelines and local microbial resistance. In the present setting (i.e. high clarithromycin resistance), despite the high observed eradication rate, sequential therapy should not be recommended (absent in guidelines, unneeded antibiotic). Bismuth-based quadruple treatment (1st, 2nd or subsequent lines) yields the highest eradication rates.Abbreviations: ALT: Altamura; BA: Bari; EGDS: esophagogastroduodenoscopy; GERD: gastro-esophageal reflux disease; H. pylori: Helicobacter pylori; ITT: intention-to-treat; PP: per-protocol; PPI: proton pump inhibitor; UBT: urea breath test.

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Molecular Assessment of Resistance to Clarithromycin in Helicobacter pylori Strains Isolated from Patients with Dyspepsia by Fluorescent In Situ Hybridization in the Center of Iran

BioMed Research International ◽

10.1155/2020/2304173 ◽

2020 ◽

Vol 2020 ◽

pp. 1-7

Author(s):

Jina Vazirzadeh ◽

Jamal Falahi ◽

Sharareh Moghim ◽

Tahmineh Narimani ◽

Rahmatollah Rafiei ◽

...

Keyword(s):

Helicobacter Pylori ◽

In Situ Hybridization ◽

Point Mutations ◽

23S Rrna ◽

Rrna Gene ◽

Clarithromycin Resistance ◽

Genotypic Analysis ◽

23S Rrna Gene ◽

H Pylori

Background and Aims. Helicobacter pylori is a common infectious bacterium mostly found in gastroduodenal diseases. The increased prevalence of clarithromycin-resistant H. pylori strains is a major challenge in the successful treatment of infections caused by this organism. The present study is aimed at detecting the clarithromycin resistance pattern of H. pylori strains isolated from gastric biopsies and evaluating point mutations of the 23S rRNA gene. Patients and methods. In the present descriptive cross-sectional study, 165 patients with gastrointestinal disorders, who were referred to the Endoscopy Center of Dr. Shariati Hospital of Isfahan, Iran, were enrolled from April to July 2018. H. pylori infection was diagnosed by culture, and susceptibility of the isolates to clarithromycin was assessed by the E-test. Minimum inhibitory concentration (MIC) values were obtained based on EUCAST recommendations. Also, fluorescence in situ hybridization (FISH) was used to determine point mutations associated with clarithromycin resistance. Results. By using culturing, H. pylori was isolated from 50.3% (83/165) gastric biopsy specimens. The overall frequency of resistance to clarithromycin was 25.3% (21/83) by the E-test. In the resistance genotypic analysis, 19 isolates had mutations. The prevalence of A2143G and A2144G mutations was 68.4% (13/19) and 31.5% (6/19), respectively. A2143C mutation was not tracked in any isolate. Two isolates with MIC>0.5 μg/mL had no mutations that could be related to other mechanisms of resistance. Conclusion. As presented in the study, the high prevalence of clarithromycin-resistant H. pylori due to point mutations of the 23S rRNA gene indicates the necessity of revising the standard treatment regimen based on antibiotic susceptibility pattern of each region.

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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.

Acta Biochimica Polonica ◽

10.18388/abp.2014_1901 ◽

2014 ◽

Vol 61 (2) ◽

Cited By ~ 22

Author(s):

Karolina Klesiewicz ◽

Paweł Nowak ◽

Elżbieta Karczewska ◽

Iwona Skiba ◽

Izabela Wojtas-Bonior ◽

...

Keyword(s):

Helicobacter Pylori ◽

Point Mutations ◽

Rflp Analysis ◽

23S Rrna ◽

Rrna Gene ◽

Efficient Management ◽

Clarithromycin Resistance ◽

Pcr Rflp ◽

23S Rrna Gene ◽

H Pylori

The occurrence of clarithromycin resistance among Helicobacter pylori strains is a major cause of the treatment failure. Resistance to this drug is conferred by point mutations in 23S rRNA gene and the most prevalent mutations are A2143G and A2142G. The aim of the study was to evaluate the occurrence of A2143G and A2142G mutations in a group of H. pylori strains resistant to clarithromycin. The study included 21 clarithromycin-resistant H. pylori strains collected between 2006 and 2009 in southern Poland. Resistance to clarithromycin was quantitatively tested with the E-test to determine the minimal inhibitory concentration (MIC value). The point mutations of H. pylori isolates were detected by PCR followed by RFLP analysis. The MIC values for clarithromycin for the analyzed strains ranged from 1.5 mg/L to 64 mg/L. Nine H. pylori strains exhibited A2143G mutation and A2142G mutation was found in 9 isolates as well. The results of RFLP analysis of 3 clarithromycin-resistant strains were negative for both mutations. The average MIC values for A2143G and A2142G mutants were 6 and 30 mg/L, respectively. Frequencies of A2143G and A2142G mutations were the same in all isolates tested. Strains with A2143G mutation exhibited lower MIC values than A2142G mutants. Application of PCR-RFLP method for detection of clarithromycin resistance allows for better and more efficient management of H. pylori infections.

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PCR-RFLP DETECTION OF POINT MUTATIONS CONFERRING RESISTANCE TO CLARITHROMYCIN OF HELICOBACTER PYLORI IN QUANG NGAI

Journal of Medicine and Pharmacy ◽

10.34071/jmp.2015.4_5.7 ◽

2015 ◽

pp. 54-61

Author(s):

Ngoc Doanh Pham ◽

Van Huy Tran ◽

Thi Minh Thi Ha

Keyword(s):

Helicobacter Pylori ◽

General Hospital ◽

Chronic Gastritis ◽

Point Mutations ◽

23S Rrna ◽

Rrna Gene ◽

Clarithromycin Resistance ◽

Urease Test ◽

Pcr Rflp ◽

H Pylori

Background: Clarithromycin resistance of H-pylori is the main cause leading to treatment failure. Aim: The purpose of this study was to determine the rate of clarithromycin resistance mutation on gene 23S ribosomal popular robonucleotide acid (rRNA) of H-pylori in patients with chronic gastritis in Quang Ngai General Hospital PCR-RFLP. Method: This is a cross-sectional study in 64 patients infected with H-pylori was determined by 3 methods and chronic gastritis proven by histology. Sample collection conducted in Quang Ngai general hospital and molecular biology tests were conducted in the medical genetics department Hue of Medical and Pharmaceutical University. Urease test, histopathological examination and perform HE staining PCR 23S rRNA gene fragment of H-pylori to determine H-pylori infection. Analysis of genetic mutations in the 23S rRNA point is performed by PCR-RFLP technique. Results: Of the 64 biopsies qualify included in the study, 41 samples with clarithromycin resistance point mutations (64%), of which 40 (62.5%) had mutations A2143A, one sample with A2142A (2%). No samples had mutations A2142C and no more than one mutation.Conclusion: This is the first time we report mutations related to clarithromycin of H-pylori in Quang Ngai province. Mutations rate is high (64%), among the common mutations, the most common mutantation is A2143G. Key words: Helicobacter pylori, clarithromycin resistance, PCR-RFLP, point mutations

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