scholarly journals PCR-Based Rapid Identification System Using Bridged Nucleic Acids for Detection of Clarithromycin-Resistant Mycobacterium avium-M. intracellulare Complex Isolates

2016 ◽  
Vol 54 (3) ◽  
pp. 699-704 ◽  
Author(s):  
Takashi Hirama ◽  
Ayako Shiono ◽  
Hiroshi Egashira ◽  
Etsuko Kishi ◽  
Koichi Hagiwara ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Detection System ◽  
Point Mutations ◽  
23S Rrna ◽  
Clinical Samples ◽  
Identification System ◽  
Rrna Gene ◽  
Clarithromycin Resistance ◽  
23S Rrna Gene ◽  
Resistant Mutants

The nontuberculous mycobacteria (NTM) cause miscellaneous disorders in humans, especially in the lungs, which present with a variety of radiological features. To date, knowledge of the pathogenic role of theMycobacterium avium-intracellularecomplex (MAC) in the human lung and the definitive criteria for initiating multidrug therapy are still lacking. However, there is little doubt that clarithromycin is the most efficacious drug among the various treatment regimens for lung NTM. In this study, with the use of a bridged nucleic acid (BNA) probe a detection system based on a real-time PCR (BNA-PCR) for the identification of the point mutations at position 2058 or 2059 in domain V of the 23S rRNA gene responsible for clarithromycin resistance was developed and has been assessed using MAC isolates from clinical samples. Out of 199 respiratory specimens, the drug susceptibility test demonstrated 12 strains resistant to clarithromycin, while the BNA-PCR showed 8 strains carrying the point mutation at position 2058 or 2059 of the 23S rRNA gene. This system revealed that there were mycobacterial strains resistant to clarithromycin which do not carry previously identified resistance genes. This paper documents a novel system for detecting clarithromycin-resistant strains and demonstrates that although these mutations are tacitly assumed to account for >90% of the reported resistant mutants, there is a significant fraction of resistant mutants that do not harbor these mutations. Therefore, unknown mechanisms affecting clarithromycin resistance remain to be elucidated.

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

scholarly journals 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

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.

scholarly journals T2182C Mutation in 23S rRNA Is Associated with Clarithromycin Resistance in Helicobacter pylori Isolates Obtained in Bangladesh

2004 ◽  
Vol 48 (9) ◽  
pp. 3567-3569 ◽  
Author(s):  
Rasel Khan ◽  
Shamsun Nahar ◽  
Jinath Sultana ◽  
Mian Mashhud Ahmad ◽  
Motiur Rahman
Keyword(s):  
Helicobacter Pylori ◽  
Gene Sequence ◽  
Point Mutations ◽  
23S Rrna ◽  
Rrna Gene ◽  
Rrna Gene Sequence ◽  
Clarithromycin Resistance ◽  
Gene Sequence Analysis ◽  
23S Rrna Gene ◽  
Transition Mutation

ABSTRACT Twelve clarithromycin-resistant (MIC, ≥1 μg/ml) Helicobacter pylori isolates were analyzed for point mutations in the 23S rRNA gene. Sequence analysis of all of the resistant isolates revealed a T-to-C transition mutation at position 2182. Transformation experiments confirmed that a single T-to-C transition mutation at position 2182 is associated with clarithromycin resistance.

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.

scholarly journals Clarithromycin resistance and 23s rrna gene point mutations of helicobacter pylori infection in children

2016 ◽  
Vol 58 (4) ◽  
pp. 371 ◽  
Author(s):  
Yeliz Çağan-Appak ◽  
Hörü Gazi ◽  
Semin Ayhan ◽  
Beyhan Cengiz-Özyurt ◽  
Semra Kurutepe ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Point Mutations ◽  
Helicobacter Pylori Infection ◽  
23S Rrna ◽  
Rrna Gene ◽  
Clarithromycin Resistance ◽  
23S Rrna Gene

Helicobacter pylori 23S rRNA gene A2142G, A2143G, T2182C, and C2195T mutations associated with clarithromycin resistance detected in Sudanese patients

2020 ◽  
Author(s):  
Aalaa Mahgoub Albasha ◽  
Maram M. Alnosh ◽  
Esraa Hassan Osman ◽  
Duha M Zeinalabdin ◽  
Amira A M Fadl ◽  
...  
Keyword(s):  
Dna Sequencing ◽  
Point Mutations ◽  
23S Rrna ◽  
Rrna Gene ◽  
Specific Pcr ◽  
Clarithromycin Resistance ◽  
23S Rrna Gene ◽  
Allele Specific ◽  
H Pylori ◽  
Allele Specific Pcr

Abstract Background: Clarithromycin resistant Helicobacter pylori (H. pylori) strains represent a worldwide health problem. These stains are usually carrying mutations within the 23S rRNA gene associated with clarithromycin resistance. This study aimed to detect H. pylori and clarithromycin resistant associated mutations from Sudanese patients with gastritis symptoms.Materials and Methods: Two hundred and eighty-eight gastric biopsies were collected using gastrointestinal endoscopy from patients with gastritis symptoms in different hospitals in Khartoum state. H. pylori was detected by PCR using primers targeting 16S rRNA and 23S rRNA. Then allele-specific PCR and DNA sequencing were used to screen for the presence of A2142G and A2143G point mutations.Results: Out of 288 samples, H. pylori was detected in 97 (33.7%) sample. Allele-specific PCR detected the variant A2142G in 9/97 (9.3%) sample, while A2143G mutation was not found in any sample. The DNA sequencing revealed the presence of mutations associated with clarithromycin-resistance in 48% (12/25) of samples; the A2142G was present in one sample, A2143G in 5 samples, T2182C in 4 samples, and C2195T in 3 samples. There was no association of 23S rRNA gene point mutations with gender, age group and geographical distribution of patients.Conclusion: This study revealed a high frequency (48%) of mutations associated with clarithromycin resistance using DNA sequencing of the 23S rRNA gene's V domain. This information should be taken into consideration before choosing optimal therapy for H. pylori eradication.

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