DETECTING POINT MUTATIONS A2142G AND A2143G IN THE 23S RRNA GENE CAUSING HELICOBACTER PYLORI RESISTANCE TO CLARITHROMYCIN BY PCR RFLP

2013 ◽  
pp. 56-63
Author(s):  
Thi Minh Thi Ha ◽  
Van Huy Tran
Keyword(s):  
Helicobacter Pylori ◽  
Point Mutations ◽  
23S Rrna ◽  
Rapid Urease Test ◽  
Rrna Gene ◽  
Peptic Ulcers ◽  
Urease Test ◽  
Pcr Rflp ◽  
Pcr Products ◽  
23S Rrna Gene

Background: Clarithromycine-resistance of H.pylori is one important cause of decreasing eradication rate of H.pylori. This study is aimed at: (1): evaluating the application of PCR RFLP in detecting point mutations A2142G and A 2143G in the 23SrRNA gene resulting Clarithromycine-resistant H.pylori. (2) determining the rate of these point mutations by PCR RFLP in patients with gastroduodenitis or peptic ulcers. Patients and methods: 38 patients with gastroduodenitis or peptic ulcer were enrolled. H.pylori infection was confirmed by rapid Urease test and PCR. PCR was performed in two phases: amplification of gene 23S rRNA containing A2143 and A2142, followed by digestion of PCR products by enzymes BbsI and BsaI in order to detecting point mutations A2143G and A2142G. Different quantities of enzyme of digestion were evaluated (5U; 7,5U; 10U; 15U and 20U) in order to determine an optimal quantity. Results: The components of digesting reaction were optimized as followings: performed in a solution volume of 20 µl, including 5 µl PCR products (to amplify gene 23S rRNA containing 2142 and 2143), 10 U enzyme (BsaI to detect A2143G, BbsI to detect A2142G), 2 µl buffer G 2X, and water for a sufficient volume. 17 point mutations A2143G were found (44.7%). Conclusion: PCR RFLP could be routinely applied to detect point mutations A2143G and A2142G in the 23S rRNA gene causing clarithromycine-resistant H.pylori. The rate of these point mutations in this group of patients was relatively high. Key words: Clarithromycin resistance, gene 23S rRNA, Helicobacter pylori

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

2014 ◽  
Vol 61 (2) ◽  
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.

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

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

STUDY ON POINT MUTATIONS AT POSITIONS 2132 AND 2143 IN 23S RRNA GENE OF HELICOBACTER PYLORI AMONG PATIENTS WITH CHRONIC GASTRITIS

2016 ◽  
pp. 12-20
Author(s):  
Thi Minh Thi Ha ◽  
Van Huy Tran ◽  
Viet Nhan Nguyen ◽  
Thanh Hoa Nguyen ◽  
Phan Tuong Quynh Le
Keyword(s):  
Helicobacter Pylori ◽  
Chronic Gastritis ◽  
Point Mutations ◽  
23S Rrna ◽  
Rrna Gene ◽  
Clarithromycin Resistance ◽  
Pcr Rflp ◽  
23S Rrna Gene ◽  
History Of ◽  
H Pylori

Background: Clarithromycin resistance in Helicobacter pylori has been found to be associated with point mutations at positions 2142 and 2143 in 23SrRNA gene. The aims of this study were: (1) to determine the rates of point mutations A2143G, A2142G and A2142C in 23SrRNA gene of H. pylori among patients with chronic gastritis by PCR-RFLP technique; and (2) to assessthe association between these mutations and some clinical, endoscopic and histopathological characteristics of chronic gastritis. Patients and methods: two hundreds and twenty six patients with H. pylori-positive chronic gastritis were determined A2143G, A2142G and A2142C mutations by PCR-RFLP technique with DNA extracted from endoscopic biopsy specimens of gastric mucosa. Results: The rate of point mutations at positions 2142 and 2143 in 23S rRNA gene of H. pylori was 35.4% in total, the A2143G and A2142G mutationsaccounted for 92.5% and 7.5% of all point mutations, respectively. No A2142C mutation was found. These mutations were not associated with age, gender,distribution of gastritis, and the presence of atrophic gastritis. The rate of A2143G mutation in groups with and without a history of clarithromycin treatment were 44.9% and 24.8%, respectively (p = 0,0065). The A2142G mutation was associated with intestinal metaplasia and/or dysplasia. Conclusion: The point mutations at positions 2142 and 2143 in 23S rRNA gene were found at a high rate in H. pylori strains amongpatients with chronic gastritis, with the absolute predominance of A2143G mutation. The A2143G mutation was associated with a history of clarithromycin treatment. Key words: 23S rRNA gene, Helicobacter pylori, A2143G, A2142G, A2142C mutation, clarithromycin resistance, chronic gastritis.

scholarly journals Helicobacter pylori 23S rRNA gene mutations associated with clarithromycin resistance in chronic gastritis in Vietnam

2018 ◽  
Vol 12 (07) ◽  
pp. 526-532 ◽  
Author(s):  
Van Huy Tran ◽  
Thi Minh Thi Ha ◽  
Phan Tuong Quynh Le ◽  
Viet Nhan Nguyen ◽  
Trung Nam Phan ◽  
...  
Keyword(s):  
Chronic Gastritis ◽  
23S Rrna ◽  
Rapid Urease Test ◽  
Rrna Gene ◽  
Wild Type ◽  
Clarithromycin Resistance ◽  
Urease Test ◽  
Pcr Rflp ◽  
23S Rrna Gene ◽  
H Pylori

Introduction: Data about the prevalence of the A2142C, A2142G, and A2143G mutations in 23S rRNA gene is still limited. The aim of this study was to determine the prevalence of these mutations in 23S rRNA gene of H. pylori vietnamese strains. Methodology: One hundred and sixty-nine patients with H. pylori-positive chronic gastritis were examined. H. pylori was detected by rapid urease test and Polymerase chain reaction (PCR). Total DNA was extracted from gastric biopsy specimens. A2142C, A2142G, and A2143G mutations were detected by DNA sequencing and PCR-restriction fragment length polymorphism (PCR-RFLP). Results: A2143G mutation was detected in 36.1% of samples, A2142G mutation in 3.6%, while A2142C mutation was not found in any case. The mixture of wild-type and mutation strains was found in 50% of specimens with A2142G, in 23% of specimens with A2143G mutation. There was no association of 23S rRNA gene point mutations with gender or age. However, an association between the heterogeneity of mutation and age was evidenced, with mean age of the group of pure A2143G higher than the group of wild-type/A2143G mixture, and rate of the wild-type/A2143G mixture higher in patients under 40 years of age. Conclusion: A2143G mutation was prominent, while A2142C mutation was not found in the 23S rRNA gene. PCR-RFLP has revealed a reliable assay allowing a rapid and cost-effective detection of clarithromycin-resistant strains. This is useful in countries as Vietnam with high prevalence of clarithromycin-resistance before choosing optimal therapy for H. pylori eradication.

The Resistance Rate of Helicobacter Pylori to Clarithromycin and Main Mutations on Bacterial Genomic Responsible for Bacterial Resistance: A Comparative Study in Children and Adults, Tehran and Iran

2019 ◽  
Vol 19 (4) ◽  
pp. 394-397 ◽  
Author(s):  
Azizollah Yousefi ◽  
Shahryar Eslami ◽  
Samileh Noorbakhsh ◽  
Morteza Haghighi ◽  
Leila TaheriNia ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Bacterial Resistance ◽  
Resistance Rate ◽  
Helicobacter Pylori Infection ◽  
23S Rrna ◽  
Antibiotics Resistance ◽  
Rapid Urease Test ◽  
Rrna Gene ◽  
Urease Test ◽  
23S Rrna Gene

Background: High resistance to common antibiotics has become a huge global dilemma in eradicating Helicobacter Pylori infection in both children and adults. The great concern is about the resistance to different classes of antibiotics especially Clarithromycin because of its widespread use. Objective: The present survey aimed to assess the resistance rate to Clarithromycin in Helicobacter Pylori isolated in patients aged less than 15 years as compared to patients older than 15 years of age. Methods: In this cross-sectional study, total 72 patients with upper gastrointestinal symptoms requiring diagnostic endoscopy referred to Rasoul-e-Akram Hospital in Tehran during one year (August 2015 to August 2016). Helicobacter Pylori infection was diagnosed in patients using the Rapid Urease Test. The antibiotics resistance was detected in genomes using the real-time polymerase chain reaction (PCR) on 23S rRNA gene. Results: In total 72 patients, 36 cases aged less than or equal to 15 years and 36 patients were older than 15 years. Of all patients in this study, 17 cases were detected with gene mutations or polymorphisms related to resistance to Clarithromycin. Overall prevalence rate of resistance was reported 23.61%. Three polymorphisms on 23S rRNA gene including A2142G, A2142C, and A2143G were revealed in 47.1%, 5.9%, and 47.1% of patients, respectively. The bacterial resistance to Clarithromycin was observed more prevalent in patients that aged older than 15 years compared to patients younger than 15 years of age. Also, frequent consumption of any type of antibiotics was significantly associated with the higher resistance of bacterium to Clarithromycin. Conclusion: The results of our study regarding the resistance of Helicobacter Pylori to Clarithromycin were similar to findings of other studies around the world. But, the Clarithromycin resistance rate was reported higher in patients older than 15 years of age and those patients who repeatedly received different types of antibiotics regardless of their age. Of all mutations in bacterial genome, the prominent mutations responsible for bacterial resistance to Clarithromycin included A2142C, A2142G, and A2143G nucleotide polymorphism on 23S rRNA gene.

scholarly journals Rapid Detection, by PCR and Reverse Hybridization, of Mutations in the Helicobacter pylori 23S rRNA Gene, Associated with Macrolide Resistance

1999 ◽  
Vol 43 (7) ◽  
pp. 1779-1782 ◽  
Author(s):  
Leen-Jan van Doorn ◽  
Yvette J. Debets-Ossenkopp ◽  
Armelle Marais ◽  
Ricardo Sanna ◽  
Francis Mégraud ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Fragment Length Polymorphism ◽  
Simultaneous Detection ◽  
Point Mutations ◽  
Macrolide Resistance ◽  
23S Rrna ◽  
Rrna Gene ◽  
Reverse Hybridization ◽  
23S Rrna Gene ◽  
H Pylori

ABSTRACT A PCR-based reverse hybridization system (research prototype kit INNO-LiPA for H. pylori resistance) was developed and evaluated for simultaneous detection of 23S ribosomal DNA point mutations, associated with macrolide resistance in Helicobacter pylori. Fifty-seven H. pylori strains (51 natural, 6 laboratory-derived artificial, 52 resistant, and 5 susceptible strains) were tested by PCR-LiPA (detecting mutations A2115→G, G2141→A, A2142→G, A2142→C, A2143→G, A2143→C, and A2143→T), DNA sequencing, restriction fragment length polymorphism, and/or hybridization to oligonucleotide probes. Results were highly concordant, but PCR-LiPA appears to be more sensitive for the simultaneous detection of multiple mutants.

Prediction of Helicobacter Pylori Clarithromycin Resistance by Detection of Point Mutations in 23S rRNA Gene

2018 ◽  
Vol 27 (3) ◽  
pp. 13-20
Author(s):  
Hayam E. Rashed ◽  
Marwa A. Mansour ◽  
Mostafa A. Soltan ◽  
Tarek I. Zahir ◽  
Yasmin A. Fahmy
Keyword(s):  
Helicobacter Pylori ◽  
Point Mutations ◽  
23S Rrna ◽  
Rrna Gene ◽  
Clarithromycin Resistance ◽  
23S Rrna Gene
Sign in / Sign up

Export Citation Format

Share Document