An Alternative, Sensitive Method to Detect <i>Helicobacter pylori</i> DNA in Feces

Horemans, Tessa; Deschacht, Maartje; Clais, Sofie; Camp, John Van; Rijk, Pim de; Holvoet, J; Assche, Tim Van; Maes, Louis; Cos, Paul

doi:10.1111/j.1523-5378.2011.00819.x

Cited by 4 publications

(3 citation statements)

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“…In H. pylori preclinical models, the evaluation focuses on the proliferation of lesions in the stomach mucosa as a result of infection, and other methods such as molecular diagnosis are also useful in the evaluation of the success of an intervention therapy [23, 24]; however, its use requires expensive equipment and is not always available. Both the low mortality rate and the chronic nature of the disease limit preclinical models of the disease.…”

Section: Discussionmentioning

confidence: 99%

Immunization with a SyntheticHelicobacter pyloriPeptide Induces Secretory IgA Antibodies and Protects Mice against Infection

Espinosa-Ramos

Caballero‐Hernández

Gómez-Flores

et al. 2019

Canadian Journal of Infectious Diseases and Medical Microbiolog

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Helicobacter pylori is a spiral Gram-negative bacterium associated with inflammation of the gastric mucosa, peptic ulcer, and gastric adenocarcinoma, whose treatment has failed due to antibiotic resistance and side effects. Furthermore, because there are no vaccines effective against H. pylori, an appropriate vaccine design targeting conserved/essential genes must be identified. In the present study, a H. pylori 50–52 kDa immunogen-derived peptide antigen with the sequence Met-Val-Thr-Leu-Ile-Asn-Asn-Glu (MVTLINNE) was used to immunize against H. pylori infection. For this, mice received an intraperitoneal injection of 100 μg of H. pylori peptide on the first week, followed by two weekly subcutaneous reinforcements and further 109 bacteria administration in the drinking water for 3 weeks. Thymic cells proliferative responses to concanavalin A, serum levels of IL-2, IL-4, IL-6, IL-10, IL-17, IFN-γ, and TNF-α cytokines, and IgG1, IgG2a, IgG2b, IgG3 IgM, and IgA immunoglobulins were evaluated. Significant (p<0.05) increases on lymphoproliferation and spleen weights after immunization were observed. In contrast, infection significantly (p<0.05) decreased lymphoproliferation, which was recovered in immunized mice. In addition, levels of serum TH1 and TH2 cytokines were not altered after immunization, except for the significant increase in IL-6 production in immunized and/or infected animals. Moreover, immunization correlated with plasma secretory IgA and IgG, whereas infection alone only elicited IgM antibodies. Peptide immunization protected 100% of mice against virulent H. pylori. MVTLINNE peptide deserves further research as an approach to the prophylaxis of H. pylori infection.

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Section: Discussionmentioning

confidence: 99%

Immunization with a SyntheticHelicobacter pyloriPeptide Induces Secretory IgA Antibodies and Protects Mice against Infection

Espinosa-Ramos

Caballero‐Hernández

Gómez-Flores

et al. 2019

Canadian Journal of Infectious Diseases and Medical Microbiolog

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“…Methods for determining LODs in spiked-stool samples were modified from [ 22 ]. To prepare spiked-stool samples, bacterial suspension of H. pylori was added to produce stool containing from 10 0 to 10 8 cells/ml.…”

Section: Methodsmentioning

confidence: 99%

Detection and genotyping of Helicobacter pylori in saliva versus stool samples from asymptomatic individuals in Northeastern Thailand reveals intra-host tissue-specific H. pylori subtypes

et al. 2018

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BackgroundTwo-thirds of the world’s population is thought to be infected by Helicobacter pylori. Although most people infected with H. pylori are asymptomatic, this pathogen is associated with several gastric pathologies including cancer. The risk factors for colonization are still unclear and the genetic diversity within individual hosts has never been clearly investigated.ResultThis study determined the prevalence of, and explored risk factors for, H. pylori infection directly from paired saliva (n = 110) and stool (n = 110) samples from asymptomatic persons in Northeast Thailand. Samples were subjected to indirect immunofluorescence assay (IFA), 16S rRNA-based real-time PCR and vacA-based semi-nested PCR. Partial vacA gene sequences of H. pylori were compared between saliva and stool samples.The overall prevalence of H. pylori infection in our asymptomatic study population was 64%. Age, gender, occupation and frequency of brushing teeth were not found to be associated with H. pylori colonization. The vacA gene was successfully sequenced from both saliva and stool samples of 12 individuals. For seven of these individuals, saliva and stool sequences fell into different clusters on a phylogenetic tree, indicating intra-host genetic variation of H. pylori.ConclusionThis study reports a high prevalence of H. pylori infection in asymptomatic persons in this region of Thailand and demonstrates that genotypes (vacA gene sequences) of H. pylori may differ between the oral cavity and intestinal tract.Electronic supplementary materialThe online version of this article (10.1186/s12866-018-1150-7) contains supplementary material, which is available to authorized users.

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“…The detection limit obtained was one bacterial cell per 100 mg of stool after heating, i.e. a 10‐fold increase in sensitivity compared with a commercially available stool DNA extraction kit [34].…”

Section: Invasive Testsmentioning

confidence: 99%

Diagnosis of Helicobacter pylori Infection

2011

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While few new methods have been proposed for Helicobacter pylori diagnosis, there are still a number of articles evaluating the current methods and trying to improve their accuracy. Invasive tests EndoscopyAttempts to diagnose Helicobacter pylori infection directly during endoscopy have been made in the past. While the observation of H. pylori per se is usually not possible, indirect evidence of its presence can be found. This year, using standard endoscopy in children, Hidaka et al. [1] were able to show that the absence of regular arrangement of collecting venules at two sites, indicated the absence of H. pylori infection with an excellent sensitivity (100%) and specificity (90%). The respective values for antral nodularity, the usual criterion, were 84% and 100%.Three studies evaluated narrow band imaging (NBI), a technique enhancing the mucosal and capillary patterns of the gastric surface. Three hundred patients were explored with conventional NBI and 5 mucosal patterns were identified, corresponding to different grades of histological gastritis [2]. Magnifying NBI was also used to investigate the changes in gastric mucosal patterns before and 12 weeks after H. pylori eradication. In patients without severe atrophy and intestinal metaplasia (IM), the changes in pit size and density of fine irregular vessels were observed with a perfect sensitivity and specificity. In contrast, no change occurred in patients with severe atrophy and IM, regardless of H. pylori eradication [3]. The authors emphasized that this method predicts histological conditions and pepsinogen levels and may be cost effective in gastric cancer surveillance [4].Confocal laser endomicroscopy, a magnifying endoscopy technique, allows an in depth analysis of the gastric mucosa. In a first study, Ji et al. [5] identified H. pylori gastritis with a 92% accuracy. The mean kappa value for inter-observer agreement was 0.78. In a second study, the same group focused on the severity of H. pylori-associated gastritis, especially atrophy and IM, with a rather good diagnostic accuracy [6]. HistologyThe ever well-known OLGA (Operative Link on Gastritis Assessment) staging system was highlighted again under the aspect of histology reporting of gastritis. It is especially valuable as it allows a prediction of the gastric cancer risk [7]. A long-term follow-up (12 years) of 93 patients confirmed that all invasive or intra-epithelial gastric neoplasia were consistently associated with high-risk (III ⁄ IV) OLGA stages [8].Immunohistochemistry can be used to assess the presence of H. pylori with more certainty. However, its systematic use in the routine of a pathology laboratory does not seem necessary because it is of no value in cases with an absence of pathological abnormality nor reactive gastropathy [6]. However, it allows the AbstractWhen an endoscopy is performed, it now becomes easier to observe indirect evidence of the presence of a Helicobacter pylori infection, given the progress of new methods including magnifying narrow band imaging or confocal l...

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An Alternative, Sensitive Method to Detect Helicobacter pylori DNA in Feces

Abstract: The proposed technique allows detection of very low quantities of H. pylori DNA in biological samples. In laboratory animal models, detailed monitoring of infection and complete clearance of infection can be demonstrated thanks to the low detection limit.

Cited by 4 publications

References 20 publications

Immunization with a SyntheticHelicobacter pyloriPeptide Induces Secretory IgA Antibodies and Protects Mice against Infection

Immunization with a SyntheticHelicobacter pyloriPeptide Induces Secretory IgA Antibodies and Protects Mice against Infection

Detection and genotyping of Helicobacter pylori in saliva versus stool samples from asymptomatic individuals in Northeastern Thailand reveals intra-host tissue-specific H. pylori subtypes

Diagnosis of Helicobacter pylori Infection

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