Development and Validation of Multiplex Quantitative PCR Assay for Detection of Helicobacter pylori and Mutations Conferring Resistance to Clarithromycin and Levofloxacin in Gastric Biopsy

Binmaeil, Hasyanee; Hanafiah, Alfizah; Rose, Isa Mohamed; Ali, Raja Affendi Raja

doi:10.2147/idr.s325056

Cited by 14 publications

(14 citation statements)

References 65 publications

(97 reference statements)

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“…Some studies have compared the agreement between the culture and the qPCR. For example, Monno et al reported an 80.6% concordance between the qPCR and the E-test for detecting clarithromycin resistance in gastric biopsies, and Bimaeil et al reported a significant concordance between these methods (Kappa = 0.85) [ 43 , 44 ]. The Maastricht V/Florence consensus report has already recognized that detecting genes and mutations associated with antibiotic resistance using molecular technologies is a valuable tool for detecting gastric biopsy specimens directly [ 14 ].…”

Section: Discussionmentioning

confidence: 99%

Helicobacter pylori Virulence Factors and Clarithromycin Resistance-Associated Mutations in Mexican Patients

et al. 2023

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Persistent infection with Helicobacter pylori (H. pylori) is an important factor in gastric diseases. The vacA and cagA virulence factors of H. pylori contribute to the development of these diseases. Triple therapy containing clarithromycin has been used to eradicate this infection. Unfortunately, resistance to this antibiotic is the primary cause of treatment failure. This study aimed to determine the prevalence of clarithromycin resistance-associated mutations and to assess the relationship between virulence factors and Mexican patients infected with H. pylori. The cagA and vacA genotypes were determined by multiplex PCR. Furthermore, a qPCR was used to identify mutations of the 23S rRNA gene. This study reported a prevalence of 84.3% of H. pylori among patients with gastric diseases, and the vacA s1m1/cagA+ genotype was the most frequent (44.8%) in antrum and corpus. Analysis of the 23S rRNA gene revealed a 19.8% prevalence of clarithromycin resistance-associated mutations. The most prevalent mutations were A2143G (56%) and A2142C (25%). A significant association (p < 0.05) between the A2142G and the vacA s1m1/cagA+ genotype was detected. In conclusion, we report a high prevalence (>15%) of clarithromycin resistance-associated mutations, and we found an association between the genotypes of virulence factors and a mutation in the 23S rRNA gene.

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Section: Discussionmentioning

confidence: 99%

Helicobacter pylori Virulence Factors and Clarithromycin Resistance-Associated Mutations in Mexican Patients

et al. 2023

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“…When drug susceptibility testing in culture is unavailable owing to time constraints, real-time PCR, 65,66 multiplex PCR, 67,68 fluorescence in situ hybridization, 69 and PCR-restriction fragment length polymorphism (PCR-RFLP) using gastric biopsy samples, 70,71 or PCR using stool samples 72,73 can be used to evaluate drug resistance to specific antimicrobial agents. Clarithromycin exerts its antibacterial activity by binding to the peptidyl transferase region of the 23S rRNA of H. pylori.…”

Section: Molecular-based Methodsmentioning

confidence: 99%

Optimizing Helicobacter pylori Treatment: An Updated Review of Empirical and Susceptibility Test-Based Treatments

et al. 2023

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As the rate of discovery of drug-resistant Helicobacter pylori cases increases worldwide, the relevant societies have updated their guidelines for primary eradication regimens. A promising strategy against drug-resistant H. pylori is tailored therapy based on the results of an antibiotic susceptibility test; however, it is difficult to apply this strategy to all cases. Although culture-based antibiotic susceptibility tests can assess resistance to any antimicrobial agent, their greatest disadvantage is the time required to draw a conclusion. In contrast, molecular-based methods, such as polymerase chain reaction, can rapidly determine the presence of resistance, although a single test can only test for one type of antimicrobial agent. Additionally, the limited availability of facilities for molecular-based methods has hindered their widespread use. Therefore, low-cost, minimally invasive, simple, and effective primary regimens are needed. Several studies have compared the efficacy of the latest primary eradication regimens against that of tailored therapies, and their results have shaped guidelines. This article reviews the latest research on empirical and tailored treatments for H. pylori infections. Evidence for the superiority of tailored therapy over empirical therapy is still limited and varies by region and treatment regimen. A network meta-analysis comparing different empirical treatment regimens showed that vonoprazan triple therapy provides a superior eradication effect. Recently, favorable results towards vonoprazan dual therapy have been reported, as it reached eradication levels similar to those of vonoprazan triple therapy. Both vonoprazan dual therapy and tailored therapy based on antibiotic susceptibility tests could contribute to future treatment strategies.

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“…137,138 At labs, and gene chip technology is emerging along with PCR technologies. [140][141][142] Compared with traditional PCR and its derivative techniques, there are multiple reaction regions on the gene chip, and oligonucleotide probes are arranged in the reaction regions to test the target genes associated with resistance to different drugs, which has significant value in the diagnosis of H. pylori infection and in detecting its antimicrobial sensitivity. 143 A study in 2020 reported that the sensitivity and specificity of gene chip detection in the diagnosis of H. pylori infection were 96.1% and 85.0%, respectively.…”

Section: Molecular Detectionmentioning

confidence: 99%

Antibiotic resistance in Helicobacter pylori: From potential biomolecular mechanisms to clinical practice

Lin

Shao

Yan

et al. 2023

Clinical Laboratory Analysis

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Helicobacter pylori (H. pylori) is a gram-negative microaerobic bacterium that is a common pathogen widely distributed around the world, especially in developing countries. 1 According to statistics from recent years, the H. pylori infection rate in developing countries exceeds 50%. 2 In West Asia, Africa, and South America, the H. pylori infection rate has been reported to be as high as 66.6%, 70.1%, and 69.4%, respectively. 3 H. pylori infection causes persistent damage to human gastric mucosa, leading to a variety of digestive

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Development and Validation of Multiplex Quantitative PCR Assay for Detection of Helicobacter pylori and Mutations Conferring Resistance to Clarithromycin and Levofloxacin in Gastric Biopsy

Cited by 14 publications

References 65 publications

Helicobacter pylori Virulence Factors and Clarithromycin Resistance-Associated Mutations in Mexican Patients

Helicobacter pylori Virulence Factors and Clarithromycin Resistance-Associated Mutations in Mexican Patients

Optimizing Helicobacter pylori Treatment: An Updated Review of Empirical and Susceptibility Test-Based Treatments

Antibiotic resistance in Helicobacter pylori: From potential biomolecular mechanisms to clinical practice

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