Application of Real-Time PCR Stool Assay for Helicobacter pylori Detection and Clarithromycin Susceptibility Testing in Brazilian Children

Scaletsky, Isabel C. A.; Aranda, Katia R. S.; Garcia, Gabriela T.; Gonçalves, Manoel Ernesto Peçanha; Cardoso, Sílvia Helena; Iriya, Kiyoshi; Silva, Neusa P.

doi:10.1111/j.1523-5378.2011.00845.x

Cited by 49 publications

(43 citation statements)

References 23 publications

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“…A further step was undoubtedly represented by the detection of point mutations conferring antibiotic resistance in bacterial DNA fecal samples. Table 1 reports the main studies on antibiotic susceptibility (clarithromycin) performed on H. pylori stool DNA by RT-PCR[32-38]. For each study, diagnostic accuracy parameter and reference standard are reported.…”

Section: Noninvasive Molecular Testsmentioning

confidence: 99%

Noninvasive molecular analysis ofHelicobacter pylori: Is it time for tailored first-line therapy?

Ierardi¹,

Giorgio²,

Iannone³

et al. 2017

WJG

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The main problem of Helicobacter pylori (H. pylori) infection management is linked to antibiotic resistances. This phenomenon has grown in the last decade, inducing a dramatic decline in conventional regimen effectiveness. The causes of resistance are point mutations in bacterial DNA, which interfere with antibiotic mechanism of action, especially clarithromycin and levofloxacin. Therefore, international guidelines have recently discouraged their use in areas with a relevant resistance percentage, suggesting first-line schedules with expected high eradication rates, i.e., bismuth containing or non-bismuth quadruple therapies. These regimens require the daily assumption of a large number of tablets. Consequently, a complete adherence is expected only in subjects who may be motivated by the presence of major disorders. However, an incomplete adherence to antibiotic therapies may lead to resistance onset, since sub-inhibitory concentrations could stimulate the selection of resistant mutants. Of note, a recent meta-analysis suggests that susceptibility tests may be more useful for the choice of first than second-line or rescue treatment. Additionally, susceptibility guided therapy has been demonstrated to be highly effective and superior to empiric treatments by both meta-analyses and recent clinical studies. Conventional susceptibility test is represented by culture and antibiogram. However, the method is not available everywhere mainly for methodology-related factors and fails to detect hetero-resistances. Polymerase chain reaction (PCR)-based, culture-free techniques on gastric biopsy samples are accurate in finding even minimal traces of genotypic resistant strains and hetero-resistant status by the identification of specific point mutations. The need for an invasive endoscopic procedure has been the most important limit to their spread. A further step has, moreover, been the detection of point mutations in bacterial DNA fecal samples. Few studies on clarithromycin susceptibility have shown an overall high sensitivity and specificity when compared with culture or PCR on gastric biopsies. On these bases, two commercial tests are now available although they have shown some controversial findings. A novel PCR method showed a full concordance between tissue and stool results in a preliminary experience. In conclusion, despite poor validation, there is increasing evidence of a potential availability of noninvasive investigations able to detect H. pylori resistances to antibiotics. These kinds of analysis are currently at a very early phase of development and caution should be paid about their clinical application. Only further studies aimed to evaluate their sensitivity and specificity will afford novel data for solid considerations. Nevertheless, noninvasive molecular tests may improve patient compliance, time/cost of infection management and therapeutic outcome. Moreover, the potential risk of a future increase of resistance to quadruple regimens as a consequence of their use on large scale and incomplete patient ad...

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Section: Noninvasive Molecular Testsmentioning

confidence: 99%

Noninvasive molecular analysis ofHelicobacter pylori: Is it time for tailored first-line therapy?

Ierardi¹,

Giorgio²,

Iannone³

et al. 2017

WJG

View full text Add to dashboard Cite

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“…Development of a detection procedure using methods such as direct sequencing may be helpful in these cases. 25,26 In a previous Japanese study, more than 90% of CAM-resistant strains of H. pylori had the A2143G mutation. 27 In this study, only one CAM-resistant strain (A2142C) was detected and a CAM resistancerelated mutation (A2143G) from fecal DNA was detected in the same patient.…”

Section: Discussionmentioning

confidence: 95%

“…Instead, in this study, we distinguished mutant‐type and wild‐type 23S rRNA sequences using software. Development of a detection procedure using methods such as direct sequencing may be helpful in these cases …”

Section: Discussionmentioning

confidence: 99%

Usefulness of detection of clarithromycin‐resistant Helicobacter pylori from fecal specimens for young adults treated with eradication therapy

et al. 2017

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PCR-based sequencing to detect CAM resistance-associated mutations using isolates or fecal samples was useful for finding antibiotic-resistant H. pylori infection. Although the specificity of the detection from fecal samples compared with antibiotic susceptibility testing was lower than that from isolates, this fecal detection method is suitable especially for asymptomatic subjects including children. Further improvement is needed before clinical application.

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“…Because of the high clinical importance of H. pylori infection, numerous diagnostic methods based on nucleic acid technology for its detection in clinical, biological, and environmental samples have been developed (22,(26)(27)(28)(29)(30)(31)(32)(33)(34).…”

Section: Discussionmentioning

confidence: 99%

Loop‐Mediated Isothermal Amplification as a Fast Noninvasive Method of Helicobacter pylori Diagnosis

Yari

Abiri

Aryan

et al. 2015

Clinical Laboratory Analysis

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Application of Real-Time PCR Stool Assay for Helicobacter pylori Detection and Clarithromycin Susceptibility Testing in Brazilian Children

Abstract: This assay proved to be appropriate for H. pylori clarithromycin susceptibility testing, particularly in children populations where a high prevalence of clarithromycin-resistant strains is suspected.

Cited by 49 publications

References 23 publications

Noninvasive molecular analysis ofHelicobacter pylori: Is it time for tailored first-line therapy?

Noninvasive molecular analysis ofHelicobacter pylori: Is it time for tailored first-line therapy?

Usefulness of detection of clarithromycin‐resistant Helicobacter pylori from fecal specimens for young adults treated with eradication therapy

Loop‐Mediated Isothermal Amplification as a Fast Noninvasive Method of Helicobacter pylori Diagnosis

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