scholarly journals Vertical Helicobacter pylori transmission from Mongolian gerbil mothers to pups

2009 ◽  
Vol 58 (5) ◽  
pp. 656-662 ◽  
Author(s):  
Ichiro Oshio ◽  
Takako Osaki ◽  
Tomoko Hanawa ◽  
Hideo Yonezawa ◽  
Cynthia Zaman ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
16S Rrna ◽  
Real Time ◽  
Mongolian Gerbil ◽  
Rt Pcr ◽  
Antibody Titres ◽  
Mother's Milk ◽  
Mother’S Milk ◽  
H Pylori

To identify the time frame and route of mother-to-child Helicobacter pylori infection, a Mongolian gerbil model was used. Four-week-old female Mongolian gerbils were infected with H. pylori, and then mated with uninfected males 2 months after infection. The offspring were sacrificed weekly after birth, and then serum, mother's milk from the stomach and gastric tissues were obtained from pups. Anti-H. pylori antibody titres were measured in sera and maternal milk using an ELISA. The stomach was cut in two in the sagittal plane, and then H. pylori colonization in mucosa was confirmed by culture and real-time RT-PCR in one specimen and by immunochemical staining in the other. Faeces and oral swabs were obtained from infected mothers, and H. pylori 16S rRNA was measured using real-time RT-PCR. H. pylori was not identified in cultures from the gastric mucosa of pups delivered by infected mothers, but H. pylori 16S rRNA was detected from 4 weeks after birth, suggesting that Mongolian gerbil pups become infected via maternal H. pylori transmission from 4 weeks of age. The anti-H. pylori antibody titre in sera of pups from infected mothers was maximum at 3 weeks of age and then rapidly decreased from 4 weeks of age. High antibody titres in mother's milk were detected during the suckling period, and GlcNAcα was detectable at 2–4 weeks of age, but disappeared as the offspring aged. Thus H. pylori seems to infect Mongolian gerbil pups from 4 weeks of age, in parallel with decreasing GlcNAcα expression in the gastric mucosa. These results suggested that H. pylori infection of Mongolian gerbil pups occurs via faecal–oral transmission from an infected mother.

scholarly journals Assessment of Helicobacter pylori Gene Expression within Mouse and Human Gastric Mucosae by Real-Time Reverse Transcriptase PCR

2001 ◽  
Vol 69 (8) ◽  
pp. 4759-4766 ◽  
Author(s):  
Bachra Rokbi ◽  
Delphine Seguin ◽  
Bruno Guy ◽  
Véronique Mazarin ◽  
Emmanuel Vidor ◽  
...  
Keyword(s):  
Gene Expression ◽  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
16S Rrna ◽  
Mouse Model ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Rt Pcr ◽  
Reverse Transcriptase Pcr ◽  
H Pylori

ABSTRACT Despite increasing knowledge on the biology of Helicobacter pylori, little is known about the expression pattern of its genome during infection. While mouse models of infection have been widely used for the screening of protective antigens, the reliability of the mouse model for gene expression analysis has not been assessed. In an attempt to address this question, we have developed a quantitative reverse transcriptase PCR (RT-PCR) that allowed the detection of minute amounts of mRNA within the gastric mucosa. The expression of four genes, 16S rRNA, ureA (encoding urease A subunit), katA (catalase), and alpA (an adhesin), was monitored during the course of a 6-month infection of mice and in biopsy samples from of 15 infected humans. We found that the selected genes were all expressed within both mouse and human infected mucosae. Moreover, the relative abundance of transcripts was the same (16S rRNA > ureA >katA > alpA), in the two models. Finally, results obtained with the mouse model suggest a negative effect of bacterial burden on the number of transcripts of each gene expressed per CFU (P < 0.05 for 16S rRNA, alpA, andkatA). Overall, this study demonstrates that real-time RT-PCR is a powerful tool for the detection and quantification ofH. pylori gene expression within the gastric mucosa and strongly indicates that mice experimentally infected with H. pylori provide a valuable model for the analysis of bacterial gene expression during infection.

scholarly journals Long-Term Infection of Mongolian Gerbils with Helicobacter pylori: Microbiological, Histopathological, and Serological Analyses

2005 ◽  
Vol 12 (2) ◽  
pp. 347-353 ◽  
Author(s):  
Shigehito Nakagawa ◽  
Takako Osaki ◽  
Yasunori Fujioka ◽  
Hiroyuki Yamaguchi ◽  
Shigeru Kamiya
Keyword(s):  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Real Time ◽  
Mononuclear Cells ◽  
Bacterial Colonization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mongolian Gerbils ◽  
Rt Pcr ◽  
H Pylori

ABSTRACT The effects of long-term infection with Helicobacter pylori on the gastric mucosa of Mongolian gerbils were examined. Colonization by H. pylori was evaluated by both microaerobic cultivation and real-time reverse transcriptase PCR (RT-PCR). Persistent infection with H. pylori in gastric mucosa was detected by real-time RT-PCR during 6 months after infection, but no H. pylori was isolated 4 months after infection by cultivation. Infiltration with neutrophils and mononuclear cells was observed from 2 months after infection. Both intestinal metaplasia and gastric atrophy were also detected from 2 months after infection. The results by enzyme-linked immunosorbent assay indicated that antibody titers against whole H. pylori antigens, H. pylori heat shock protein 60 (HSP60), and Escherichia coli GroEL were significantly higher in the infected gerbils than in noninfected gerbils. After long-term infection with H. pylori for 18 months, marked atrophy of gastric mucosa and multiple cysts in the submucosa were observed in the glandular stomach of the infected gerbils. In addition, squamous cell papilloma with hyperkeratosis was observed in cardia of all the infected gerbils. These results indicate that evaluation of bacterial colonization during long-term infection can be done by real-time RT-PCR and that mucosal damage might be induced by host immune response against whole H. pylori antigen.

scholarly journals Direct detection of Helicobacter pylori from biopsies of patients in Lagos, Nigeria using real-time PCR—a pilot study

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
A. Ajayi ◽  
T. Jolaiya ◽  
S. I. Smith
Keyword(s):  
Helicobacter Pylori ◽  
16S Rrna ◽  
Real Time ◽  
Real Time Pcr ◽  
Direct Detection ◽  
Low Cost ◽  
Treatment Options ◽  
Combined Cycle ◽  
Diagnostic Methods ◽  
H Pylori

Abstract Objective Prompt diagnosis of Helicobacter pylori infection is essential for proper treatment and eradication of the pathogen because prolonged infection could lead to gastric cancer. Sensitive and cost effective diagnostic methods are key to guiding treatment options that will reduce mortality. This study was aimed at detecting H. pylori from biopsies of peptic ulcer patients. Real-time PCR using TaqMan and EvaGreen assays targeting 16S rRNA and ureA genes were used to detect H. pylori DNA extracted from 40 biopsy samples comprising 20 biopsies obtained from the antrum and 20 from the corpus of 20 patients undergoing endoscopy for duodenal ulcer investigation in Lagos, Nigeria. Results H. pylori was detected in 80% of the biopsy samples by combined cycle threshold (Ct) and melting temperature (Tm) values. Mean Ct value for ureA gene ranged from 21.40 to 37.53 and 22.71 to 35.44 for 16SrRNA gene. Average melting temperatures (Tm) of 81.57 and 82.90 °C among amplicons of ureA and 16S rRNA were observed respectively. H. pylori DNA was generally detected in biopsies collected from antrum and corpus. Real-time PCR in the diagnosis of H. pylori can be considered a simple, low cost and efficient alternative or addition to the gold standard.

Detection of Helicobacter pylori in gastric cancer tissue through histopathology, immunohistochemistry and real-time reverse transcription-PCR

2020 ◽  
Vol 15 (12) ◽  
pp. 1131-1137
Author(s):  
Carlos A Castaneda ◽  
Miluska Castillo ◽  
Joselyn Sanchez ◽  
Sandro Casavilca ◽  
Juvenal Sanchez ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Helicobacter Pylori ◽  
Real Time ◽  
Gastric Cancer Tissue ◽  
Cancer Tissue ◽  
Kappa Index ◽  
Rt Pcr ◽  
Reverse Transcription Pcr ◽  
Gene Copies ◽  
H Pylori

Aim: Helicobacter pylori is usually detected based on hematoxylin–eosin (H–E) features, but, immunohistochemistry (IHC) and real-time PCR (RT-PCR) are more precise in chronic-gastritis. We evaluated the relevance of these tests in Peruvian gastric cancer samples. Materials & methods: We performed and evaluated H–E, IHC staining and RT-PCR in 288 gastric tumors. Slides were independently evaluated by three pathologists. Results: H. pylori was detected in 167/287 through H–E, 140/288 through IHC and 175/288 through RT-PCR, and positive-status were associated (p < 0.001). H. pylori detection by H–E had a good concordance with IHC (kappa index = 0.632) but poor with RT-PCR (kappa index = 0.317). Higher median gene-copies were found in high H. pylori density through H–E or IHC (p < 0.001). Conclusion: H–E evaluation is accurate in gastric cancer, and IHC and RT-PCR can complement its results.

scholarly journals Real-Time PCR Screening for 16S rRNA Mutations Associated with Resistance to Tetracycline in Helicobacter pylori

2005 ◽  
Vol 49 (8) ◽  
pp. 3166-3170 ◽  
Author(s):  
Erik Glocker ◽  
Marco Berning ◽  
Monique M. Gerrits ◽  
Johannes G. Kusters ◽  
Manfred Kist
Keyword(s):  
Helicobacter Pylori ◽  
16S Rrna ◽  
Real Time ◽  
Real Time Pcr ◽  
Tetracycline Resistance ◽  
16S Rrna Genes ◽  
Rrna Genes ◽  
Rrna Gene ◽  
Pcr Screening ◽  
H Pylori

ABSTRACT The effectiveness of recommended first-line therapies for Helicobacter pylori infections is decreasing due to the occurrence of resistance to metronidazole and/or clarithromycin. Quadruple therapies, which include tetracycline and a bismuth salt, are useful alternative regimens. However, resistance to tetracycline, mainly caused by mutations in the 16S rRNA genes (rrnA and rrnB) affecting nucleotides 926 to 928, are already emerging and can impair the efficacies of such second-line regimens. Here, we describe a novel real-time PCR for the detection of 16S rRNA gene mutations associated with tetracycline resistance. Our PCR method was able to distinguish between wild-type strains and resistant strains exhibiting single-, double, or triple-base-pair mutations. The method was applicable both to DNA extracted from pure cultures and to DNA extracted from fresh or frozen H. pylori-infected gastric biopsy samples. We therefore conclude that this real-time PCR is an excellent method for determination of H. pylori tetracycline resistance even when live bacteria are no longer available.

scholarly journals Activation of IκB Kinase β and NF-κB Is Essential for Helicobacter pylori-Induced Chronic Gastritis in Mongolian Gerbils

2007 ◽  
Vol 76 (2) ◽  
pp. 781-787 ◽  
Author(s):  
Ayako Yanai ◽  
Shin Maeda ◽  
Wataru Shibata ◽  
Yohko Hikiba ◽  
Kei Sakamoto ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Mongolian Gerbil ◽  
Proinflammatory Cytokine ◽  
Mongolian Gerbils ◽  
Activated Macrophages ◽  
Macrophage Depletion ◽  
Iκb Kinase ◽  
H Pylori

ABSTRACT The Mongolian gerbil model of Helicobacter pylori infection resembles human gastritis. In this study, we investigated the role of NF-κB activation in H. pylori-infected gerbils. Activated macrophages were significantly increased in H. pylori-infected gastric mucosa and were identified as being important cells with potent activation of NF-κB, which plays an important part in producing proinflammatory cytokines. Macrophage depletion by the administration of clodronate resulted in milder inflammation in gerbils infected with H. pylori. In macrophages, the inhibition of IκB kinase β (IKKβ), which is a critical kinase for NF-κB activation, resulted in lower proinflammatory cytokine expression caused by heat-killed H. pylori cells. Furthermore, treatment with IKKβ inhibitor resulted in milder inflammation in gerbils with H. pylori gastritis. Collectively, our data suggest that H. pylori-mediated gastric inflammation critically depends on the efficient recruitment and activation of macrophages, with sufficient NF-κB activation.

scholarly journals Expression of Melatonin Synthesizing Enzymes inHelicobacter pyloriInfected Gastric Mucosa

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
Cezary Chojnacki ◽  
Tomasz Popławski ◽  
Janusz Blasiak ◽  
Jan Chojnacki ◽  
Russel J. Reiter ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Mrna Expression ◽  
Normal Level ◽  
Breath Test ◽  
Histological Analysis ◽  
Urea Breath Test ◽  
Unknown Etiology ◽  
Rt Pcr ◽  
H Pylori

Helicobacter pyloricolonization of gastric mucosa causes pain of unknown etiology in about 15–20% of infected subjects. The aim of the present work was to determine the level of expression of enzymes involved in the synthesis of melatonin in gastric mucosa of asymptomatic and symptomaticH. pyloriinfected patients. To diagnoseH. pyloriinfection, histological analysis and the urea breath test (UBT C13) were performed. The levels of mRNA expression of arylalkylamine-N-acetyltransferase (AA-NAT) and acetylserotonin methyltransferase (ASMT) were estimated in gastric mucosa with RT-PCR. The level of AA-NAT expression and AMST was decreased inH. pyloriinfected patients and was increased afterH. pylorieradication. We conclude that decreased expression of melatonin synthesizing enzymes, AA-NAT and ASMT, in patients with symptomaticH. pyloriinfection returns to normal level afterH. pylorieradication.

scholarly journals Prevalence and genotyping of Helicobacter pylori in endoscopic biopsy samples from a Chinese population

2019 ◽  
Vol 43 (1) ◽  
pp. 21-28
Author(s):  
Hao Yu ◽  
Yingjia Mao ◽  
Lijie Cong ◽  
Zhiyong Wang ◽  
Hua Zhang ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Real Time ◽  
Chinese Population ◽  
Rflp Analysis ◽  
Endoscopic Biopsy ◽  
Gastric Biopsy ◽  
Rapid Urease Test ◽  
Urease Test ◽  
H Pylori

Abstract Background: Helicobacter pylori inhabit the gastric mucosa of humans and are associated with several gastrointestinal diseases which include gastric cancer, peptic ulcer, chronic gastritis and gastric mucosa-associated lymphoid tissue lymphoma. Helicobacter pylori exhibit a high degree of genetic variability and are associated with its epidemiological, pathological characteristics and dynamics of transmission. The objective of the study was to determine the prevalence and genetic heterogeneity of H. pylori isolated from endoscopic biopsy samples from a Chinese population. Methods: Gastric biopsy samples from 86 patients (males, 55; females, 35) who presented to the endoscopic section for various gastrointestinal abnormalities were collected. The samples were subjected to a real-time polymerase chain reaction (PCR) and microbial culture for the isolation of H. pylori. Further, the isolates were subjected to randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) analysis. Results: Of the 86 gastric biopsy samples, 61 (70.9%) samples were positive for rapid urease test and 37 (43%) samples (28 from male and nine from female) grew H. pylori. Among the biopsy samples subjected to real-time PCR, 39 (45.3%) samples were found to be positive for H. pylori. The RAPD analysis yielded 15 different patterns (four to 17 different sized fragments per strain). The phylogenetic analysis of RAPD yielded 22 clusters at a similarity level ranging from 63% to 100%. RFLP analysis yielded nine different patterns (two to six different sized fragments per strain). Two major restriction patterns were identified, of which 14 (37.8%) strains forms the most common pattern (genotype I) followed by five (13.5%, genotype II) strains with an intra-strain similarity of 100%. Conclusions: The overall prevalence of H. pylori was 45.3%. Despite reports on the declining trend in the prevalence of H. pylori infections, our prevalence rate was still higher than those reported from other developed countries. However, further studies involving a large sample size and covering more regions of China is highly warranted.

Prevalence and genotyping of Helicobacter pylori in endoscopic biopsy samples from a Chinese population

2018 ◽  
Vol 0 (0) ◽  
Author(s):  
Hao Yu ◽  
Yingjia Mao ◽  
Lijie Cong ◽  
Zhiyong Wang ◽  
Hua Zhang ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Real Time ◽  
Chinese Population ◽  
Rflp Analysis ◽  
Endoscopic Biopsy ◽  
Gastric Biopsy ◽  
Rapid Urease Test ◽  
Urease Test ◽  
H Pylori

AbstractBackground:Helicobacter pylori inhabit the gastric mucosa of humans and are associated with several gastrointestinal diseases which include gastric cancer, peptic ulcer, chronic gastritis and gastric mucosa-associated lymphoid tissue lymphoma. Helicobacter pylori exhibit a high degree of genetic variability and are associated with its epidemiological, pathological characteristics and dynamics of transmission. The objective of the study was to determine the prevalence and genetic heterogeneity of H. pylori isolated from endoscopic biopsy samples from a Chinese population.Methods:Gastric biopsy samples from 86 patients (males, 55; females, 35) who presented to the endoscopic section for various gastrointestinal abnormalities were collected. The samples were subjected to a real-time polymerase chain reaction (PCR) and microbial culture for the isolation of H. pylori. Further, the isolates were subjected to randomly amplified polymorphic DNA (RAPD) and restriction fragment length polymorphism (RFLP) analysis.Results:Of the 86 gastric biopsy samples, 61 (70.9%) samples were positive for rapid urease test and 37 (43%) samples (28 from male and nine from female) grew H. pylori. Among the biopsy samples subjected to real-time PCR, 39 (45.3%) samples were found to be positive for H. pylori. The RAPD analysis yielded 15 different patterns (four to 17 different sized fragments per strain). The phylogenetic analysis of RAPD yielded 22 clusters at a similarity level ranging from 63% to 100%. RFLP analysis yielded nine different patterns (two to six different sized fragments per strain). Two major restriction patterns were identified, of which 14 (37.8%) strains forms the most common pattern (genotype I) followed by five (13.5%, genotype II) strains with an intra-strain similarity of 100%.Conclusions:The overall prevalence of H. pylori was 45.3%. Despite reports on the declining trend in the prevalence of H. pylori infections, our prevalence rate was still higher than those reported from other developed countries. However, further studies involving a large sample size and covering more regions of China is highly warranted.

scholarly journals Development of an easy-to-use urease kit for detecting Helicobacter pylori in canine gastric mucosa

2021 ◽  
pp. 1977-1987
Author(s):  
Chularat Hlaoperm ◽  
Kiattawee Choowongkomon ◽  
Chantima Pruksakorn ◽  
Jatuporn Rattanasrisomporn
Keyword(s):  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Real Time ◽  
Real Time Pcr ◽  
Color Change ◽  
Sybr Green ◽  
The Other ◽  
Urease Enzyme ◽  
H Pylori ◽  
Commercial Kit

Background and Aim: Helicobacter pylori is an important pathogen in humans and animals involved in chronic gastritis, leading to the development of gastric cancer. Urease produced by H. pylori is an enzyme that promotes bacterial colonization and can be used clinically as a biomarker of H. pylori infection as part of a rapid urease test (RUT). A test with high specificity (95-100%) would be more convenient and faster than histopathology, bacterial culture, and polymerase chain reaction (PCR). The aim of this study was to develop a simple, cheap, and fast kit for detecting H. pylori infection in the gastric mucosa of canines, which can be used in clinical practice for diagnosing infection with this bacterium. Materials and Methods: The RUT assays developed were prepared using 1% agar, 1% sodium phosphate monobasic, and 1% urea followed by the addition of 3% methyl red indicator. The cutoff value of sensitivity of the RUT assay was established using the urease of H. pylori ATCC 43504 and color change was monitored for 24 h. Comparisons of the sensitivity to H. pylori ATCC 43504 were made between the developed RUT assays and the Hp Fast™ commercial kit. Then, the limit of detection for H. pylori ATCC 43504 number was analyzed by the SYBR Green real-time PCR assay to measure the copy number of the ureC gene. Gastric biopsy samples from the antrum, body, and fundus of the stomach were collected from eight canines presenting with vomiting and gastroenteritis. Analyses were performed on fresh samples using the developed RUT assays and the Hp Fast™ commercial kit, which were read within 24 h; then, the results were confirmed with SYBR Green real-time PCR. The specificity of the RUT assays was tested with a number of different bacteria, including Staphylococcus pseudintermedius, Proteus spp., Pseudomonas aeruginosa, Klebsiella pneumoniae, Enterococcus spp., Escherichia coli, and Salmonella spp.; H. pylori ATCC 43504 was used as a positive control. Results: The results showed that the developed assays were sensitive to the urease enzyme at 0.1 mg/mL. The lowest detection limit of this assay for H. pylori ATCC 43504 was found to be 102 copies at 30 min. The sensitivity of detection of H. pylori in gastric biopsies of canines occurred in a minimum of 30 min. The RUT showed similar results to the Hp Fast™ commercial kit. In the developed RUT, the color change of the test from red to yellow could be clearly distinguished between the color of the positive test and the negative one; however, in the commercial Hp Fast™, it was difficult to observe the gel color change in the negative pH range of 5.8 and the positive pH of 6.5. The developed RUT was specific for H. pylori and did not detect any of the other tested bacteria. The test kit can also be stored for 6 months at 4°C. Conclusion: The sensitivity of the developed assays allowed the detection of urease enzyme at a minimum concentration of 0.1 mg/mL. Our RUT could also detect H. pylori from one in eight canine specimens at a minimum of 102 copies within 30 min. This RUT is specific to H. pylori as it did not detect any of the other tested bacteria.

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