Helicobacter pylori Mutations Conferring Resistance to Fluoroquinolones and Clarithromycin among Dyspeptic Patients Attending a Tertiary Hospital, Tanzania (original) (raw)

Molecular Detection of Clarithromycin and Fluoroquinolones Resistance in Helicobacter Pylori Infection, Directly Applied to Gastric Biopsies, in an Urban Brazilian Population

Arquivos de Gastroenterologia, 2016

Background - Antimicrobial resistance is the major factor leading to eradication failure in H. pylori treatment. Molecular tests are useful to detect genetic mutations predictive of clarithromycin and fluoroquinolones resistance. Knowledge of the local prevalence rate of resistance is important to define the best recommended treatment. Objective - To assess the prevalence of primary resistance of H. pylori to clarithromycin and fluoroquinolones, using a molecular test, in a Southeastern urban Brazilian population. Methods - A total of 72 H. pylori seropositive patients [65% female, mean age 39 (19-73) years] never treated before for this infection were studied. All patients underwent gastroscopy in addition to antrum and corpus biopsies and molecular test GenoType HelicoDR (Hain Life Science, Germany) to detect H. pylori and point mutations in genes responsible for clarithromycin and fluoroquinolone resistance. The molecular procedure was divided into three steps: DNA extraction fro...

Helicobacter pylori from Peptic Ulcer Patients in Uganda Is Highly Resistant to Clarithromycin and Fluoroquinolones: Results of the GenoType HelicoDR Test Directly Applied on Stool

BioMed Research International

Background. Around 70–90% of peptic ulcer disease (PUD) is due to Helicobacter pylori and requires treatment with antimicrobials to which these bacteria are susceptible. Common H. pylori diagnostic tests do not provide drug susceptibility data. Using the GenoType HelicoDR PCR test designed for gastric biopsies for simultaneous detection of H. pylori and its resistance to clarithromycin (CLA)/fluoroquinolones (FLQ), we present evidence for stool as an optional test specimen and also provide data on prevalence of H. pylori resistance to CLA and FLQ in Uganda. Methods. Stool from 142 symptomatic PUD patients at three hospitals in Kampala was screened for H. pylori using a rapid antigen test. The GenoType HelicoDR test was run on all H. pylori antigen positives to determine PCR positivity and resistance to CLA/FLQ. Results. Thirty-one samples (22%) were H. pylori antigen positive, and 21 (68%) of these were H. pylori PCR positive. Six of the 21 (29%) were resistant to CLA and eight to F...

Mutational Analysis of Clarithromycin and Levofloxacin Resistance in Helicobacter pylori from Gastric Biopsy Specimens in a Tertiary Care Hospital in Dhaka, Bangladesh

Bangladesh Journal of Medical Microbiology, 2019

Background: Clarithromycin and Levofloxacin are most frequently included in the standard triple therapies for H. pylori eradication in our country. Resistance to clarithromycin and fluoroquinolones are particularly related with treatment failure. Objectives: The objective of this study was to detect, clarithromycin and levofloxacin resistance associated with gene mutations in H. pylori directly from gastric biopsies using an allele specific primer-PCR (ASP-PCR) assay. Materials and Methods: Gastric biopsy specimens were collected from 143 adult dyspeptic patients, from Department of Gastroenterology, BSMMU and Dhaka Medical College Hospital (DMCH), during the period of March, 2018 to February, 2019. H. pylori was identified by rapid urease test, ureC gene by PCR, histological staining and culture. ASP-PCR was used to identify 23S rRNA gene and gyrA gene mutation predictive of clarithromycin and levofloxacin resistant H. pylori respectively. Results: H. pylori positive cases were 32....

Low Helicobacter pylori primary resistance to clarithromycin in gastric biopsy specimens from dyspeptic patients of a city in the interior of São Paulo, Brazil

2013

Background: Clarithromycin, amoxicillin, and a pump proton inhibitor are the most common drugs recommended as first-line triple therapy for H.pylori treatment, which results in eradication rates close to 80%, varying regionally, principally due to emergency cases and increases of clarithromycin resistant strains. Nucleotide substitutions at the H. pylori domain V of the 23S rRNA fraction are involved in the macrolide resistance and the A2142G and A2143G mutations are predominant in clinical isolates worldwide including in Brazil. As H. pylori culture is fastidious, we investigated the primary occurrence of H. pylori A2142G and A2143G rDNA 23S mutations using a molecular approach directly on gastric biopsies of dyspeptic patients consecutively attended at Hospital das Clinicas of Marilia, São Paulo, Brazil.

Macrolide and fluoroquinolone resistance in Helicobacter pylori isolates: an experience at a tertiary care centre in Pakistan

JPMA. The Journal of the Pakistan Medical Association, 2012

To assess fluoroquinolone and clarithromycin susceptibility pattern along with the types of genomic mutations involved in the resistance of Helicobacter pylori isolates. The cross-sectional study was conducted at the Department of Pathology and Microbiology, Aga Khan University Hospital, Karachi, from June 2009 to July 2010, and comprised 162 gastric biopsy samples which were tested with GenoTypeHelicoDR (Hain Lifescience GmbH, Germany), a reverse hybridisation multiplex polymerase chain reaction (PCR) line probe assay (LiPA). Also, 23S rRNA (ribosomal ribonucleic acid) gene was analysed with three-point mutations at A2146G, A2146C and A2147G for clarithromycin, and gyrA gene was analyzed at two codon positions 87 and 91 for fluoroquinolone susceptibility testing. SPSS 19 was used for statistical analyses. Clarithromycin resistance was seen in 60 (37.0%) of the isolates mainly involving mutation at A2147G (85%) followed by A2146G (n=35; 21.6%) and A2146C (n=19; 11.6%). Fluoroquinolo...

Prevalence of single nucleotide mutation in clarithromycin resistant gene of Helicobacter pylori: a 32-months prospective study by using hybridization real time polymerase chain reaction

Journal of the Medical Association of Thailand = Chotmaihet thangphaet, 2012

Polymerase chain reaction (PCR) for detection of single nucleotide mutations in 23S rRNA gene of clarithromycin resistance Helicobacter pylori were investigated worldwide. In Thailand, the prevalence of these mutations had not been extensively investigated. The authors conducted a 32-months prospective study to estimate the prevalence of clarithromycin resistant Helicobacter pylori and to compare the sensitivity and specificity of hybridization real time PCR for 23S rRNA gene of Helicobacter pylori detection with the combination of rapid urease test, immunohistochemistry straining and Helicobacter pylori culture. A total of 200 patients with endoscopic examination with gastric biopsy were enrolled from January 2006 to September 2008. Eight gastric specimens were biopsied and performed rapid urease test, immunohistochemistry straining for Helicobacter pylori, Helicobacter pylori culture and hybridization real time PCR for 23S rRNA dectection, as well as single nucleotide mutation det...

Helicobacter pylori resistance to clarithromycin and fluoroquinolones in a pediatric population in Turkey: A cross‐sectional study

Helicobacter

Background: Helicobacter pylori antimicrobial resistance is gradually increasing around the world. However, there are a limited number of studies reporting on this issue in the pediatric population. In this study, we aimed to determine H pylori resistance to clarithromycin and fluoroquinolones in the pediatric patients living in Kırıkkale province that were detected with H pylori in gastric biopsies. Moreover, we also aimed to investigate the concordance between the histopathologic and molecular methods used in the diagnosis of H pylori infection. Materials and Methods: Patients aged 2-18 years who had a history of epigastric pain and/or nausea persisting for longer than 1 month underwent upper gastrointestinal endoscopy. Biopsies were taken from the gastric antral mucosa. In the samples detected with H pylori in the histopathologic examination, the presence of H pylori and H pylori resistance to clarithromycin and fluoroquinolones was investigated using the GenoType HelicoDR test which allows the detection of wild-type and mutant genes. The strains detected with more than one mutant gene are defined as hetero-resistant strains. Results: The 93 patients that underwent DNA extraction and amplification included 68 (73.1%) girls and 25 (26.9%) boys with a median age of 15 ± 2.62 (range 6-17) years. The overall concordance for the diagnosis of H pylori infection between histopathology and PCR was 94%, and H pylori resistance to clarithromycin and fluoroquinolones was 27% and 15%, respectively. Conclusions: The high H pylori resistance to clarithromycin and fluoroquinolones among the pediatric patients in our region implicates that the antibiotic sensitivity of strains should be studied prior to administration in accordance with the recommendations provided in the guidelines. Moreover, the presence of hetero-resistant strains in our patients may be a reason for treatment failure.

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

2020

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 muta...