INDEL-typing: a new method of differentiation of Helicobacter pylori strains

Bacteriology ◽  
2020 ◽  
Vol 5 (1) ◽  
pp. 8-13
Author(s):  
V.M. Sorokin ◽  
◽  
A.S. Vodop’janov ◽  
R.V. Pisanov ◽  
◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Geographical Origin ◽  
Diversity Index ◽  
Minimal Spanning Tree ◽  
Typing Method ◽  
Indel Markers ◽  
H Pylori ◽  
First Time ◽  
Tree Method ◽  
High Diversity

For the first time INDEL-typing method of Helicobacter pylori strains is offered. Analysis of the nucleotide sequences of 69 strains from the local database was carried out. Ten INDEL-markers were identified, five of which, with the highest variability, served as the basis for the differentiation scheme. INDEL-typing of twenty one strains of H. pylori of different geographical origin was carried out. Аnd fifteen individual genotypes with a high diversity index were identified (DI = 0.95). For cluster analysis, the MST (minimal spanning tree) method was used, which demonstrated a clear distribution of clusters according to the geographical origin of the strains studied. In addition to the proposed method of typing, if necessary, the MLVAtyping method can be used. Keywords: Helicobacter pylori, PCR, INDEL, MST dendrogram, MLVA, geographical origin

scholarly journals Geographical diversity of Helicobacter pylori strains circulating in the European Part of the Russian Federation

2021 ◽  
Vol 11 (4) ◽  
pp. 701-706
Author(s):  
V. M. Sorokin ◽  
A. V. Svarval ◽  
A. S. Vodop'janov ◽  
R. V. Pisanov
Keyword(s):  
Russian Federation ◽  
Geographical Origin ◽  
Diversity Index ◽  
European Part ◽  
Typing Method ◽  
Population Total ◽  
Saint Petersburg ◽  
Indel Markers ◽  
The Russian Federation ◽  
H Pylori

The aim of the study was to identify INDEL markers and study geographical origin of regional H. pylori strains circulating in the European part of the Russian Federation. The study included 56 strains of H. pylori isolated in three regions of the Russian Federation: Saint Petersburg, Astrakhan and Rostov Regions. Genomic DNA was isolated using a set of Probe NA (DNA Technology, Russia), according to the manufacturer’s instructions. Detection of INDEL markers hp5605, hp6405, hp340, hp1390, hp3660 was performed using PCR. Clustering of the identified INDEL genotypes and building a phylogenetic tree were performed using the BioNumerics 7.6 and GrapeTree software packages. 21 strains from the GenBank database with known geographical origin were used as reference strains. In 20 strains from Saint Petersburg, 13 individual genotypes were identified, while 17 strains belong to the European cluster (hpEurope), 2 strains belong to the hspEAsia cluster and one strain belongs to the hspWAfrica cluster. The most common genotype identified in the European cluster includes six strains from Saint Petersburg and two strains from the GenBank database. For further differentiation of these strains, the VNTR typing method was used, which allowed identifying eight individual genotypes in eight strains. Fifty-six studied russian strains are represented by thirty individual genotypes, which reflects the high heterogeneity of strains circulating in the European part of the Russian Federation. The most frequent genotype is represented by two hpEurope strains, one strain from the Astrakhan region, as well as 5 and 6 strains from the Rostov Region and Saint Petersburg, respectively. The vast majority of Russian strains (52/56) belong to the hpEurope population, while two strains from Saint Petersburg are included in the hspEAsia population, and one strain from Saint Petersburg and the Astrakhan Region is included in the hspWAfrica population. Total, 77 H. pylori strains are represented by 37 individual genotypes with a high diversity index (DI = 0.95), which allows us to consider the proposed INDEL typing method as an independent method for genotyping H. pylori strains. Taking into consideration the complexity of the problem of accurately determining the geographical origin of H. pylori strains, the proposed simple and convenient method of INDEL typing of H. pylori strains, based on an available PCR method becomes very relevant and allows us to conduct an adequate primary analysis of the geographical origin of Russian H. pylori strains.

scholarly journals Correlation between Quantitative 13C-Urea Breath Test and Helicobacter pylori Treatment Success in a Population-Based Cohort

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Doron Boltin ◽  
Zohar Levi ◽  
Tsachi Tsadok Perets ◽  
Hemda Schmilovitz-Weiss ◽  
Rachel Gingold-Belfer ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Breath Test ◽  
Treatment Success ◽  
Population Based ◽  
Urea Breath Test ◽  
First Line Therapy ◽  
13C Urea Breath Test ◽  
Helicobacter Pylori Treatment ◽  
H Pylori ◽  
First Time

Background. There are continual efforts to identify factors which influence the success of first-line therapy for Helicobacter pylori (H. pylori) infection. The 13C-urea breath test result (C13-UBT) utilizes H. pylori urease activity and is a highly accurate diagnostic assay. We aimed to determine whether the magnitude of C13-UBT result is related to treatment success. Methods. Adult patients who underwent a first-time 13C-urea breath test between January 2010 and January 2016 were included. In order to isolate a naïve test-and-treat population who were unlikely to have undergone an initial endoscopy-based H. pylori test, we excluded patients > 45 years and those with a previous C13-UBT. Data were extracted from the Clalit Health Services laboratory database. Results. A total of 94,590 subjects (36.1% male, age 28.5 ± 6.0 years) who underwent a first-time C13-UBT during the study period were included. C13-UBT was positive in 48,509 (51.3%) subjects. A confirmatory posttreatment C13-UBT was performed in 18,375 (37.8%), and eradication was successful in 12,018 (65.4%). The mean C13-UBT recording was 20.6 ± 16.2 DOB in subjects with successful eradication and 19.5 ± 13.1 DOB in subjects with treatment failure (OR, 1.01; 95% CI 1.00-1.01, p<0.01). Among patients in the upper quintile of C13-UBT measurement, eradication was achieved in 67.6%, compared to 62.6% in the lower quintile (OR, 1.22; 95% CI 1.11-1.35, p<0.01). Subjects in the top 1 percentile (C13-UBT ≥ 70 DOB) achieved eradication in 75.0%, compared to 65.3% among subjects with C13-UBT < 70 DOB (OR, 1.59; 95% CI 1.05-2.41, p<0.01). Conclusions. The superiority in H. pylori eradication observed in subjects with a higher C13-UBT DOB is small but significant. Further studies should examine the physiological and microbiological basis for this finding.

scholarly journals Functional Analysis of the Helicobacter pylori cag Pathogenicity Island Reveals Both VirD4-CagA-Dependent and VirD4-CagA-Independent Mechanisms

2002 ◽  
Vol 70 (2) ◽  
pp. 665-671 ◽  
Author(s):  
Matthias Selbach ◽  
Stefan Moese ◽  
Thomas F. Meyer ◽  
Steffen Backert
Keyword(s):  
Helicobacter Pylori ◽  
Gene Knockout ◽  
Shuttle Vector ◽  
Pathogenicity Island ◽  
Type Iv Secretion ◽  
Host Responses ◽  
Type Iv ◽  
Knockout Mutants ◽  
H Pylori ◽  
First Time

ABSTRACT The type IV secretion machinery encoded by the cag pathogenicity island (PAI) of Helicobacter pylori has been implicated in a series of host responses during infection. Here, we analyzed the function of 12 cag PAI genes from both cag I and cag II loci, including the complete virB/D complex (virB4, virB7, virB8, virB9, virB10, virB11, and virD4). We monitored interleukin-8 (IL-8) secretion, CagA translocation and tyrosine phosphorylation, and induction of a scattering (“hummingbird”) phenotype upon H. pylori infection of AGS gastric epithelial cells. For the first time, we have complemented individual cag PAI gene knockout mutants with their intact genes expressed from a shuttle vector and showed that complemented CagA and VirD4 restored wild-type function. Our results demonstrate that phenotypic changes and phosphorylation of CagA depended on all virB/D genes and several other genes of the cag PAI. Induction of IL-8 secretion depended largely on the same set of genes but was independent of CagA and VirD4. Thus, CagA translocation and induction of IL-8 secretion are regulated by VirD4-CagA-dependent and VirD4-CagA-independent mechanisms, respectively. The function of VirD4 as a possible adapter protein which guides CagA into the type IV secretion channel is presented in a model.

scholarly journals Oral rinses in growth inhibition and treatment of Helicobacter pylori infection

2020 ◽  
Author(s):  
Dharmendra Kashyap ◽  
Budhadev Baral ◽  
Tarun Prakash Verma ◽  
Charu Sonkar ◽  
Debi Chatterji ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Oral Cavity ◽  
Growth Inhibition ◽  
Growth Pattern ◽  
Gastric Biopsy ◽  
Striking Difference ◽  
Ags Cells ◽  
Oral Rinse ◽  
H Pylori ◽  
First Time

Abstract Background Helicobacter pylori ( H. pylori ) is well-known for its role in chronic gastritis and gastric cancer. Eradication of these carcinogenic bacteria from the gut is one of the challenges for clinicians. The complexity of treatment mainly owes to antibiotic resistance and relapse due to an additional reservoir in the oral cavity. Our study emphases the isolation of H. pylori from distinct habitats of the gut microenvironment (gastric biopsy and gastric juice) and its subsequent characterization. We have also evaluated the effect of various oral rinses on isolated H. pylori from different anatomical locations of included subjects. Results The possible strains isolated from two different habitats of the same subject shows a striking difference in their growth pattern. Promisingly, some of the included oral rinses are efficient in growth inhibition as per recommended 30 sec treatment. The subsequent evaluation shows that oral rinse B (among A-E) is most effective and down-regulates the expression of one of the potent H. pylori gene, CagA, in the infected gastric adenocarcinoma (AGS) cells. Conclusion Our study, for the first time, revealed that H. pylori, isolated from the different habitat of the same subject, show a different growth pattern. The expression of H. pylori pathogenic gene (CagA) was down-regulated by the use of oral rinses. Hence, oral rinses will reduce the H. pylori in the oral cavity and help to control its migration from oral to the gastric compartment and may be used as an adjuvant treatment option for its re-infection.

scholarly journals Oral rinses in growth inhibition and treatment of Helicobacter pylori infection

2020 ◽  
Author(s):  
Dharmendra Kashyap ◽  
Budhadev Baral ◽  
Tarun Prakash Verma ◽  
Charu Sonkar ◽  
Debi Chatterji ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Oral Cavity ◽  
Growth Inhibition ◽  
Growth Pattern ◽  
Gastric Biopsy ◽  
Striking Difference ◽  
Ags Cells ◽  
Oral Rinse ◽  
H Pylori ◽  
First Time

Abstract Background Helicobacter pylori ( H. pylori ) is well-known for its role in chronic gastritis and gastric cancer. Eradication of these carcinogenic bacteria from the gut is one of the challenges for clinicians. The complexity of treatment mainly owes to antibiotic resistance and relapse due to an additional reservoir in the oral cavity. Our study emphases the isolation of H. pylori from distinct habitats of the gut microenvironment (gastric biopsy and gastric juice) and its subsequent characterization. We have also evaluated the effect of various oral rinses on isolated H. pylori from different anatomical locations of included subjects. Results The possible strains isolated from two different habitats of the same subject shows a striking difference in their growth pattern. Promisingly, some of the included oral rinses are efficient in growth inhibition as per recommended 30 sec treatment. The subsequent evaluation shows that oral rinse B (among A-E) is most effective and down-regulates the expression of one of the potent H. pylori gene, CagA, in the infected gastric adenocarcinoma (AGS) cells. Conclusion Our study, for the first time, revealed that H. pylori, isolated from the different habitat of the same subject, show a different growth pattern. The expression of H. pylori pathogenic gene (CagA) was down-regulated by the use of oral rinses. Hence, oral rinses will reduce the H. pylori in the oral cavity and help to control its migration from oral to the gastric compartment and may be used as an adjuvant treatment option for its re-infection.

scholarly journals Implications for Induction of Autoimmunity via Activation of B-1 Cells by Helicobacter pylori Urease

2006 ◽  
Vol 74 (1) ◽  
pp. 248-256 ◽  
Author(s):  
Shingo Yamanishi ◽  
Tadasu Iizumi ◽  
Eri Watanabe ◽  
Masumi Shimizu ◽  
Shigeru Kamiya ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
B Cells ◽  
Immunoglobulin A ◽  
Polymyxin B ◽  
Autoimmune Disorders ◽  
Innate Immune Responses ◽  
Autoreactive Antibodies ◽  
H Pylori ◽  
First Time

ABSTRACT Besides various gastroduodenal diseases, Helicobacter pylori infection may be involved in autoimmune disorders like rheumatoid arthritis (RA) or idiopathic thrombocytopenic purpura. Such autoimmune disorders are often associated with autoreactive antibodies produced by B-1 cells, a subpopulation of B lymphocytes. These B-1 cells are mainly located in the pleural cavity or mucosal compartment. The existence of H. pylori urease-specific immunoglobulin A (IgA)-producing B cells in the mucosal compartment and of their specific IgM in the sera of acutely infected volunteers suggests the possibility that urease stimulates mucosal innate immune responses. Here, we show for the first time that purified H. pylori urease predominantly stimulates the B-1-cell population rather than B-2 cells, which produce antigen-specific conventional antibodies among splenic B220+ B cells. The fact that such stimulation of B-1 cells was not affected by the addition of polymyxin B indicates that the effect of purified H. pylori urease was not due to the contamination with bacterial lipopolysaccharide. Furthermore, the production of various B-1-cell-related autoreactive antibodies such as IgM-type rheumatoid factor, anti-single-stranded DNA antibody, and anti-phosphatidyl choline antibody was observed when the splenic B cells were stimulated with purified H. pylori urease in vitro. These findings suggest that H. pylori components, urease in particular, may be among the environmental triggars that initiate various autoimmune diseases via producing autoreactive antibodies through the activation of B-1 cells. The findings shown here offer important new insights into the pathogenesis of autoimmune disorders related to H. pylori infection.

scholarly journals Helicobacter pylori: getting to grips with the guidance

2020 ◽  
pp. flgastro-2020-101571
Author(s):  
David I F Wands ◽  
Emad M El-Omar ◽  
Richard Hansen
Keyword(s):  
Helicobacter Pylori ◽  
Treatment Options ◽  
Ulcer Disease ◽  
Treatment Regimens ◽  
The North ◽  
Paediatric Practice ◽  
Global Increase ◽  
American Society ◽  
H Pylori ◽  
First Time

Helicobacter pylori is a Gram-negative bacterium that inhabits the mucus layer above the gastric mucosa. While infection rates vary by region, the global prevalence is estimated at 50%. While asymptomatic carriage is common, infection can result in significant morbidity and mortality from complications including peptic ulcer disease, atrophic gastritis and gastric cancer. Paediatric and adult practices diverge due to differences in complication rate, symptomatology, practicalities with investigations and treatment options. Widespread use of standard antibiotic regimens has however resulted in a rapid global increase in antibiotic resistance and treatment failure in all ages. There is urgent need to optimise treatment regimens and maximise first-time eradication rates. This need is reflected in the latest guidelines from the European Society for Paediatric Gastroenterology Hepatology and Nutrition and the North American Society for Pediatric Gastroenterology, Hepatology and Nutrition for paediatric practice and the Maastricht Guidelines for adult practice. This article aims to provide a practical overview of the investigations and management of H. pylori by comparing and contrasting these guidelines.

scholarly journals Oral rinses in growth inhibition and treatment of Helicobacter pylori infection

2020 ◽  
Author(s):  
Dharmendra Kashyap ◽  
Budhadev Baral ◽  
Tarun Prakash Verma ◽  
Charu Sonkar ◽  
Debi Chatterji ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Oral Cavity ◽  
Growth Inhibition ◽  
Growth Pattern ◽  
Gastric Biopsy ◽  
Striking Difference ◽  
Oral Rinse ◽  
H Pylori ◽  
First Time ◽  
Pathogenic Gene

Abstract Background Helicobacter pylori ( H. pylori ) is well-known for its role in chronic gastritis and gastric cancer. Eradication of these carcinogenic bacteria from the gut is one of the challenges for clinicians. The complexity of treatment mainly owes to antibiotic resistance and relapse due to an additional reservoir in the oral cavity. Our study emphases the isolation of H. pylori from distinct habitats of the gut microenvironment (gastric biopsy and gastric juice) and its subsequent characterization. We have also evaluated the effect of various oral rinses on isolated H. pylori from different anatomical locations of included subjects.Results The possible strains isolated from two different habitats of the same subject shows a striking difference in their growth pattern. Promisingly, some of the included oral rinses are efficient in growth inhibition as per recommended 30 sec treatment. The subsequent evaluation shows that oral rinse B (among A-E) is most effective and down-regulates the expression of one of the potent H. pylori gene, CagA, in the infected gastric adenocarcinoma (AGS) cells.Conclusion Our study, for the first time, revealed that H. pylori, isolated from the different habitat of the same subject, show a different growth pattern. The expression of H. pylori pathogenic gene (CagA) was down-regulated by the use of oral rinses. Hence, oral rinses will reduce the H. pylori in the oral cavity and help to control its migration from oral to the gastric compartment and may be used as an adjuvant treatment option for its re-infection.

scholarly journals Helicobacter Pylori Different Virulence Genes and the Risk of Gastric Cancer in Iranian Patients

2020 ◽  
Author(s):  
Maryam Kianmehr ◽  
Ahmad Hormati ◽  
Mohsen Zargar ◽  
Roohollah Fateh ◽  
Razieh Nazari
Keyword(s):  
Gastric Cancer ◽  
Helicobacter Pylori ◽  
Virulence Genes ◽  
Control Group ◽  
Gastric Diseases ◽  
Qrt Pcr ◽  
Digestive Diseases ◽  
H Pylori ◽  
High Diversity ◽  
Iranian Patients

Abstract Background: Some disease-specific Helicobacter pylori virulence genes can be used for predicting the outcome of diseases. Thus, the current study aimed to explore the frequency of the vacA, cagA, sabA, dupA, babA, oipA, and iceA1 genes in H. pylori isolates, and to determine whether any association exists between the expression of these genes and gastric cancer (GC).Methods: H. pylori isolates were collected from individuals with digestive diseases. The cagA, vacA, dupA, sabA, babA, oipA, and iceA1 genes were determined by PCR. The qRT-PCR was used for expression analysis of the tested genes. Results: The presence of the cagA, vacA, dupA, sabA, babA, oipA and iceA1 genes in H. pylori isolates were 41.3%, 95.5%, 31.0%, 93.1%, 55.1%, 82.7%, and 62.0%, respectively. The cagA, dupA, and babA expression in the GC patients was statistically higher than that of the control group (P <0.05). Conclusions: Our results indicated a high diversity and frequency of H. pylori virulence genes among individuals with gastric diseases in the Iranian population. The cagA, dupA, and babA expression were significantly higher in the GC patients, thus, it may suggest that screening of these genes may help identify peoples at higher risk for GC.

scholarly journals NF-κB Activation during Acute Helicobacter pylori Infection in Mice

2007 ◽  
Vol 76 (2) ◽  
pp. 551-561 ◽  
Author(s):  
Richard L. Ferrero ◽  
Patrick Avé ◽  
Delphine Ndiaye ◽  
Jean-Christophe Bambou ◽  
Michel R. Huerre ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Transgenic Mice ◽  
Nuclear Factor Κb ◽  
Helicobacter Felis ◽  
Cell Responses ◽  
Mucosal Cells ◽  
H Pylori ◽  
First Time

ABSTRACT Nuclear factor κB (NF-κB) plays a key regulatory role in host cell responses to Helicobacter pylori infection in humans. Although mice are routinely used as a model to study H. pylori pathogenesis, the role of NF-κB in murine cell responses to helicobacters has not been studied in detail. We thus investigated the abilities of different Helicobacter isolates to induce NF-κB-dependent responses in murine gastric epithelial cells (GECs) and in transgenic mice harboring an NF-κB-responsive lacZ reporter gene. H. pylori and Helicobacter felis strains up-regulated the synthesis in mouse GECs of the NF-κB-dependent chemokines KC (CXCL1) and MIP-2 (CXCL2). These responses were cag pathogenicity island (cagPAI) independent and could be abolished by pretreatment with a pharmacological inhibitor of NF-κB. Consistent with the in vitro data, experimental Helicobacter infection of transgenic mice resulted in increased numbers of GECs with nuclear β-galactosidase activity, which is indicative of specific NF-κB activation. The numbers of β-galactosidase-positive cells in mice were significantly increased at day 1 postinoculation with wild-type H. pylori strains harboring or not harboring a functional cagPAI, compared to naive animals (P = 0.007 and P = 0.04, respectively). Strikingly, however, no differences were observed in the levels of gastric NF-κB activation at day 1 postinoculation with H. felis or at day 30 or 135 postinoculation with H. pylori. This work demonstrates for the first time the induction of NF-κB activation within gastric mucosal cells during acute H. pylori infection. Furthermore, the data suggest that helicobacters may be able to regulate NF-κB signaling during chronic infection.

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