Use of Immunoblot Assay To Define Serum Antibody Patterns Associated with Helicobacter pylori Infection and with H. pylori-Related Ulcers

Serology has been used worldwide to detect Helicobacter pylori infection. Using an immunoblot assay with an antigen from strain ATCC 43579, we sought to determine the antibodies which were good markers of colonization and the antibody patterns associated with ulcers or atrophy. Out of 98 dyspeptic patients, 41 were colonized byH. pylori, based on a positive culture or on positive results of both a urease test and direct examination. These 41 patients were seropositive by an enzyme immunoassay, and 12 of them had ulcers and 29 had evidence of atrophy. Fifty-seven of the 98 patients were noncolonized. Twenty-five of the 57 had evidence of gastric atrophy, and 10 were seropositive; 5 of these 10 had ulcers. By Western blot analysis, 12 antibodies were significantly more frequent in sera from colonized patients, and they produced immunoreactive bands at 125, 87, 74, 66, 54, 48, 46, 42, 35, 30, 16 and 14 kDa. The presence of at least one band at 54, 35, or 42 kDa was the best marker of infection (sensitivity, 95%; specificity, 82%). In the group of colonized patients, none of the antibody patterns were correlated to gastric atrophy. Conversely, the presence of a band at 125, 87, or 35 kDa was statistically associated with the presence of an ulcer. The simultaneous presence of bands at 87 and 35 kDa predicted the risk of ulcers with 83% sensitivity and 69% specificity. By using CagA-positive and VacA-positive strains and CagA-negative and VacA-negative isogenic mutants, the antigens corresponding to the bands at 125 and 87 kDa were shown to be CagA and VacA, respectively. On the other hand, the 35-kDa antigen is a novel uncharacterized component ofH. pylori. These results may help to optimize the composition of antigenic preparations for serologic detection ofH. pylori colonization. Immunoblot assay would be useful for screening patients at high risk of ulcers.

Download Full-text

Precancerous gastric conditions in high Helicobacter pylori prevalence areas: comparison between Eastern European (Lithuanian, Latvian) and Asian (Taiwanese) patients

Medicina ◽

10.3390/medicina43080080 ◽

2007 ◽

Vol 43 (8) ◽

pp. 623 ◽

Cited By ~ 3

Author(s):

Laimas Jonaitis ◽

Audrius Ivanauskas ◽

Dainius Jančiauskas ◽

Konrads Funka ◽

Agnese Sudraba ◽

...

Keyword(s):

Helicobacter Pylori ◽

Intestinal Metaplasia ◽

The Elderly ◽

Gastric Atrophy ◽

Eastern European ◽

Urease Test ◽

Gastric Biopsies ◽

The Mean ◽

H Pylori ◽

Positive Results

The aim of the study was to compare the prevalence and severity of precancerous condition – gastric atrophy and intestinal metaplasia (IM) between Eastern European (Lithuania and Latvia) and Asian (Taiwan) countries in population older than 55 years. Methods. Patients aged 55 years and older, referred for upper endoscopy due to dyspeptic symptoms, were included in the study. Gastric biopsies were histological investigated according modified Sydney classification. Helicobacter pylori (H. pylori) was detected if any two of three methods (urease test, histology, and serology) were positive. Results. Overall 322 patients included: 52 from Taiwan (TW), 171 from Latvia (LV) and 99 from Lithuania (LT). There were 227 (70%) females and 95 (30%) males. The mean age of TW patients was significantly lower (61.0±5.8 years), than of LV (68.1±7.3 years) and LT (66.5±7.5 years) patients. H. pylori was established in 224 (69.6%) patients. H. pylori positivity was established in 43 (82.7%) TW patients, in 112 (65.5%) LV patients, and in 69 (69.7%) LT patients (P>0.05). In H. pylori-infected patients, any atrophy either in the corpus or in the antrum of the stomach was detected in 26 (60.5%) TW patients, in 40 (35.7%) LV patients, and in 36 (52.2%) LT patients (between TW and LV patients P<0.005). Severe atrophy (grade 2 or 3) detected in 8 (18.6%) TW patients, in 17 (15.2%) LV patients, and in 18 (26.1%) LT patients (P>0.05). Intestinal metaplasia was detected in 22 (51.2%) TW patients, in 37 (33.0%) LV patients and in 31 (44.9%) LT patients among countries (P>0.05). There were no significant differences in proportions of different degrees of both atrophy and intestinal metaplasia among countries. Intestinal metaplasia was found in 79 (77.5%) of 102 patients with any degree of atrophy and in 11 (9.0%) of 122 patients without atrophy (P<0.0001). We found strong statistically significant correlations between atrophy and intestinal metaplasia in antrum (r=0.89), P<0.01, and corpus (r= 0.73), P<0.01. Conclusions. The prevalence of H. pylori in the elderly population is still high in LT, LV, and TW. There are no significant differences in prevalence of gastric atrophy and intestinal metaplasia among TW, LT, and LV. There is a strong correlation between gastric atrophy and intestinal metaplasia.

Download Full-text

Prevalence of Helicobacter pylori vacA, cagA, cagE1, cagE2, dupA and oipA Genotypes in Patients With Gastrointestinal Diseases

ACTA MEDICA IRANICA ◽

10.18502/acta.v58i7.4417 ◽

2020 ◽

Author(s):

Hossein Masoumi Asl ◽

Ali Badamchi ◽

Shima Javadinia ◽

Siamak Khaleghi ◽

Leila Tehraninia ◽

...

Keyword(s):

Gastric Cancer ◽

Helicobacter Pylori ◽

Gastrointestinal Diseases ◽

Cancer Group ◽

Urease Test ◽

High Prevalence ◽

Vaca Gene ◽

H Pylori ◽

Positive Results ◽

Gastroenterology Unit

Helicobacter pylori (H. pylori) is a bacterium that resides in the human stomach, which is associated with gastroduodenal diseases. We investigate the prevalence of cagA, vacA, oipA, cagE1, cagE2 and dupA genotypes in H. pylori isolated from patients with Gastric ulcer, duodenal ulcer, and Gastric Cancer. Collected 74 samples from the Gastroenterology Unit of the Rasool Akram Hospital were included in this study. Gastric disorders were identified by endoscopy .gastric cancer was further confirmed by histopathology. H. pylori were detected by the urease test. Subsequently, DNA was extracted from gastric tissue of the subjects with the CLO-test yielded positive results. In general, 74 patients with a mean age of 53.45 years (Range 22 to 86-year-old), including 45 men and 29 women, were studied. Among 74 H. pylori-positive patients, 70 (94.5%) patients were positive for the cagA gene. About 95.8% (23/24) of the patients with gastric carcinoma were dupA positive and VacA gene (91.8%). The oipA genotype was detected in 71 (96%) of H.pylori positive samples. This gene was more common in patients with gastritis rather than cancer group. Also, 97.2% of 74 H. pylori isolates were cagE2-positive. In 25 patients with PUD, the occurrence percent of cagA+/VacA+, cagA+/Vac- , cagA- /VacA+ and cagA- /VaxA- genotypes were found 80%, 12%, 4.2% and 4.2 respectively. The results of the present study suggest that a high prevalence of virulent factors could contribute to the risk of developing gastroduodenal diseases.

Download Full-text

Is the Remarkable Contradiction between Histology and 14C Urea Breath Test in the Detection of Helicobacter pylori due to False-negative Histology or False-positive 14C Urea Breath Test?

Journal of International Medical Research ◽

10.1177/147323000503300604 ◽

2005 ◽

Vol 33 (6) ◽

pp. 632-640 ◽

Cited By ~ 10

Author(s):

AK Gurbuz ◽

AM Ozel ◽

Y Narin ◽

Y Yazgan ◽

H Baloglu ◽

...

Keyword(s):

Helicobacter Pylori ◽

False Positive ◽

Breath Test ◽

False Negative ◽

Urea Breath Test ◽

Predictive Values ◽

Urease Test ◽

H Pylori ◽

Micro Organisms ◽

Positive Results

We assessed the diagnostic value of the 14C urea breath test (UBT) in the detection of Helicobacter pylori compared with histology and the rapid urease test (RUT). The study included 68 patients (22 men and 46 women) with dyspeptic symptoms. H. pylori status was evaluated by 14C UBT, RUT and histology. Sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy were determined for 14C UBT and for RUT. Histology revealed dense yeast-like micro-organisms in the biopsy specimens in all patients with false-positive results by 14C UBT (n = 8), a significantly higher proportion than in patients with negative 14C UBT (five of 31). The low specificity of the H. pylori14C UBT should not be neglected by accepting histology results as false-negative. Gastric mucosal colonization by yeast-like micro-organisms with urease activity can account for the high frequency of false-positive results for 14C UBT.

Download Full-text

Serodiagnosis and Bacterial Genome of Helicobacter pylori Infection

Toxins ◽

10.3390/toxins13070467 ◽

2021 ◽

Vol 13 (7) ◽

pp. 467

Author(s):

Aina Ichihara ◽

Hinako Ojima ◽

Kazuyoshi Gotoh ◽

Osamu Matsushita ◽

Susumu Take ◽

...

Keyword(s):

Helicobacter Pylori ◽

Antibody Titer ◽

Bacterial Genome ◽

Serum Antibody ◽

Gene Mutations ◽

Bacterial Genomes ◽

Western Blots ◽

A Genome ◽

Vaca Gene ◽

H Pylori

The infection caused by Helicobacter pylori is associated with several diseases, including gastric cancer. Several methods for the diagnosis of H. pylori infection exist, including endoscopy, the urea breath test, and the fecal antigen test, which is the serum antibody titer test that is often used since it is a simple and highly sensitive test. In this context, this study aims to find the association between different antibody reactivities and the organization of bacterial genomes. Next-generation sequences were performed to determine the genome sequences of four strains of antigens with different reactivity. The search was performed on the common genes, with the homology analysis conducted using a genome ring and dot plot analysis. The two antigens of the highly reactive strains showed a high gene homology, and Western blots for CagA and VacA also showed high expression levels of proteins. In the poorly responsive antigen strains, it was found that the inversion occurred around the vacA gene in the genome. The structure of bacterial genomes might contribute to the poor reactivity exhibited by the antibodies of patients. In the future, an accurate serodiagnosis could be performed by using a strain with few gene mutations of the antigen used for the antibody titer test of H. pylori.

Download Full-text

PREVALENCE OF INFECTION WITH CAGA-POSITIVE HELICOBACTER PYLORI STRAINS AMONG CHILDREN AND ADOLESCENTS IN SOUTHERN BRAZIL

Arquivos de Gastroenterologia ◽

10.1590/s0004-28032014000300003 ◽

2014 ◽

Vol 51 (3) ◽

pp. 180-185 ◽

Cited By ~ 9

Author(s):

Juliana Ghisleni de OLIVEIRA ◽

Cristina Helena Targa FERREIRA ◽

Anna Carolina Saraiva CAMERIN ◽

Cláudia Augustin ROTA ◽

Luíse MEURER ◽

...

Keyword(s):

Helicobacter Pylori ◽

Children And Adolescents ◽

Polymerase Chain Reaction Method ◽

Southern Brazil ◽

Worldwide Distribution ◽

Clinical Presentations ◽

Urease Test ◽

Polymerase Chain ◽

H Pylori ◽

Low Prevalence

Context Helicobacter pylori (H. pylori) has a worldwide distribution, but the prevalence of infection, virulence factors, and clinical presentation vary widely according to the studied population. In Brazil, a continental country composed of several ethnicities and cultural habits, the behavior of infection also appears to vary, as many other studies have shown. Objectives Describe the prevalence of infection with cagA-positive H. pylori strains in a group of children and adolescents who underwent esophagogastroduodenoscopy in Porto Alegre, Rio Grande do Sul. Methods Fifty-four gastric biopsy specimens of children and adolescents with H. pylori infection demonstrated by histology, urease test and molecular analysis were tested for the presence of cagA positive H. pylori strains by the polymerase chain reaction method. Results he prevalence of cagA-positive H. pylori was 29.6% (95% confidence interval, 18 to 43.6%). There were no statistically significant differences in clinical or demographic characteristics or in the endoscopic and histological features of patients infected with cagA-positive strains as compared with those infected by cagA-negative strains. Conclusions he study showed a low prevalence of infection with cagA-positive H. pylori strains among children and adolescents who underwent EGD in southern Brazil, in comparison to studies conducted with children from other regions of Brazil. There was no association between the presence of cagA-positive strains and more severe clinical presentations in the studied sample.

Download Full-text

Frequency of rdx A Deletion Mutation Conferring Metronidazole Resistance in Helicobacter pylori

10.51429/ejmm29308 ◽

2020 ◽

Vol 29 (3) ◽

pp. 59-64

Author(s):

Hanaa M. El Maghraby ◽

Samar Mohaseb

Keyword(s):

Helicobacter Pylori ◽

Epigastric Pain ◽

Deletion Mutation ◽

Rapid Urease Test ◽

Rrna Gene ◽

University Hospitals ◽

Metronidazole Resistance ◽

Urease Test ◽

Gastric Biopsies ◽

H Pylori

Background: Metronidazole is one of the antimicrobial drugs that can be used in combination with other drugs for eradication of Helicobacter pylori (H. pylori).Unfortunately, metronidazole resistance in H. plori is an increasing health problem which may be attributed to inactivation of many genes as rdx A gene. Objective: To determine the frequency of rdx A deletion mutation in H. pylori detected in infected patients attending at the Gastroenterology Unit, Zagazig University Hospitals. Methodology: Two gastric biopsies were taken from each enrolled patient by endoscopy. H.pylori detection was done by rapid urease test and polymerase chain reaction (PCR) amplification of 16S rRNA gene. Deletion mutation in rdx A gene was detected by conventional PCR. Results: Out of 134 doubled gastric biopsies obtained from 134 patients, 52.2% were positive for H. pylori. Epigastric pain, vomiting and gastritis were significantly associated with detection of H. pylori infection (p˂ 0.05). Deletion mutation of rdx A gene was detected in 28.6% of H. pylori positive specimens obtained from infected patients. Conclusion: Deletion mutation of rdx A gene is a frequent determinant of rdx A inactivation conferring metronidazole resistance among H. pylori.

Download Full-text

Incidence of Gastric Metaplasia and Helicobacter pylori Infection in Duodenal Bulb – With Specific Reference to Patients With Duodenal Ulcers

Diagnostic and Therapeutic Endoscopy ◽

10.1155/dte.6.17 ◽

1999 ◽

Vol 6 (1) ◽

pp. 17-23 ◽

Cited By ~ 4

Author(s):

Minoru Kawaguchi ◽

Toshihiko Saito

Keyword(s):

Helicobacter Pylori ◽

Duodenal Bulb ◽

Mucosal Injury ◽

Transitional Zone ◽

Specific Reference ◽

Rapid Urease Test ◽

Duodenal Ulcers ◽

Gastric Metaplasia ◽

Urease Test ◽

H Pylori

We determined the incidence of gastric metaplasia in the duodenal bulb of duodenal ulcer patients and the Helicobacter pylori (H. pylori) infection rate at sites with gastric metaplasia. Biopsy of the duodenal bulb showed the presence of gastric metaplasia in 61 of 86 patients (71%) overall and in 18 of 47 patients (38.3%) who had gastrectomy at an early gastric cancer. The histological diagnosis of H. pylori infection showed good agreement (83.3%) with the result of the rapid urease test, indicating that H. pylori occurs in regions with gastric metaplasia. This finding suggests that H. pylori infects gastric metaplasia in the duodenal bulb, causing mucosal injury, which is then transformed into duodenal ulcers. The exact mechanism by which gastric metaplasia is caused is unknown, but it is believed to occur in the transitional zone in the duodenal mucosa.

Download Full-text

Can probiotics improve efficiency and safety profile of triple Helicobacter pylori eradication therapy? A prospective randomized study

Vojnosanitetski pregled ◽

10.2298/vsp150415127g ◽

2016 ◽

Vol 73 (11) ◽

pp. 1044-1049 ◽

Cited By ~ 3

Author(s):

Sasa Grgov ◽

Tomislav Tasic ◽

Biljana Radovanovic-Dinic ◽

Daniela Benedeto-Stojanov

Keyword(s):

Helicobacter Pylori ◽

Randomized Study ◽

Eradication Therapy ◽

Saccharomyces Boulardii ◽

Rapid Urease Test ◽

Group I ◽

Urease Test ◽

Probiotic Strains ◽

The Difference ◽

H Pylori

Background/Aim. Some studies suggest the benefit of applying different probiotic strains in combination with antibiotics in the eradication of Helicobacter pylori (H. pylori) infection. The aim of this study was to evaluate the effect of co-administration of multiple probiotic strains with triple H. pylori eradication therapy. Methods. This prospective study included 167 patients with dyspeptic symptoms and chronic gastritis who were diagnosed with H. pylori infection and randomized into two groups. The group I of 77 patients underwent triple eradication therapy, for 7 days, with lansoprazole, 2 ? 30 mg half an hour before the meal, amoxicillin 2 ? 1.000 mg per 12 hours and clarithromycin 2 ? 500 mg per 12 hours. After the 7th day of the therapy, lansoprazole continued at a dose of 30 mg for half an hour before breakfast for 4 weeks. The group II of 90 patients received the same treatment as the patients of the group I, with the addition of the probiotic cultures in the form of a capsule comprising Lactobacillus Rosell-52, Lactobacillus Rosell-11, Bifidobacterium Rosell-1755 and Saccharomyces boulardii, since the beginning of eradication for 4 weeks. Eradication of H. pylori infection control was performed 8 weeks after the therapy by rapid urease test and histopathologic evaluation of endoscopic biopsies or by stool antigen test for H. pylori. Results. Eradication of H. pylori infection was achieved in 93.3% of the patients who received probiotics with eradication therapy and in 81.8% of patients who were only on eradication therapy without probiotics. The difference in eradication success was statistically significant, (p < 0.05). The incidence of adverse effects of eradication therapy was higher in the group of patients who were not on probiotic (28.6%) than in the group that received probiotic (17.7%), but the difference was not statistically significant. Conclusion. Multiple probiotic strains addition to triple eradication therapy of H. pylori achieves a significantly better eradication success, with fewer side effects of antibiotics.

Download Full-text

Antibiotic resistance of Helicobacter pylori in the European part of the Russian Federation: first results

Terapevticheskii arkhiv ◽

10.26442/00403660.2020.08.000761 ◽

2020 ◽

Vol 92 (8) ◽

pp. 24-28

Author(s):

I. V. Maev ◽

D. N. Andreev ◽

V. M. Govorun ◽

E. N. Ilina ◽

Yu. A. Kucheryavyy ◽

...

Keyword(s):

Helicobacter Pylori ◽

Antibiotic Resistance ◽

Russian Federation ◽

European Part ◽

Diffusion Method ◽

Rapid Urease Test ◽

Urease Test ◽

First Results ◽

The Russian Federation ◽

H Pylori

Aim. Determine the primary antibiotic resistance of Helicobacter pylori (H. pylori) strains isolated from patients living in the European part of the Russian Federation. Materials and methods. As part of a clinical laboratory study, from 2015 to 2018, 27 gastrobiopsy samples obtained from H. pylori-infected patients were analyzed. H. pylori infection was verified using a rapid urease test or a 13C-urea breath test. The values of the minimum inhibitory concentration (MIC) of antibiotics were determined by the diffusion method using E-test strips (BioMerieux, France) according to the recommendations of the manufacturer. The sensitivity of the isolates was determined for 6 antibacterial drugs (amoxicillin, clarithromycin, metronidazole, levofloxacin, tetracycline, rifampicin). Results. According to the data obtained, resistance to amoxicillin was 0%, clarithromycin 11.1%, metronidazole 59.3%, levofloxacin 3.7%, tetracycline 0%, and rifampicin 14.8%. Dual resistance to clarithromycin and metronidazole was recorded in two isolates (7.4%). Conclusion. Thus, the first results of the evaluation of H. pylori antibiotic resistance in the European part of the Russian Federation indicate a low resistance of the microorganism to clarithromycin and quite high to metronidazole.

Download Full-text

Association between gallbladder diseases and Helicobacter pylori infection

AL-Kindy College Medical Journal ◽

10.47723/kcmj.v16i2.263 ◽

2020 ◽

Vol 16 (2) ◽

pp. 30-34

Author(s):

Ali Hamid Abd-Almahdi ◽

Zuhair B. Kamal

Keyword(s):

Helicobacter Pylori ◽

Urban Areas ◽

Weight Status ◽

Gallbladder Disease ◽

Case Series ◽

Rapid Urease Test ◽

Gallbladder Diseases ◽

Case Series Study ◽

Urease Test ◽

H Pylori

Background: Helicobacter pylorus is one of the most harmful human pathogens & carcinogen. Of the world's population, more than 50% has H. pylori in their upper gastrointestinal tracts. It has been linked to a variety of extra gastric disorders. In correlation to hepatobiliary diseases; recently, the bacterium has been implicated as a risk factor for various diseases ranging from chronic cholecystitis and primary biliary sclerosing cholangitis to gall bladder cancer and primary hepatic carcinomas. However, the association between Helicobacter pylori (H. pylori) and gallbladder diseases is still vague and is controversial. Aim of study: To elucidate the association of H pylori and gallbladder diseases (calculus, acalculous, polyp), the feasibility of using rapid urease test in post-operative diagnosis, and many factors related bacterium. Subjects & methods: This case series study was conducted in Al-Kindy Teaching hospital - surgical unit during a period extended for 2 years from September 2016 to September 2018, where patients suffered from signs and symptoms of gallbladder disease were interviewed using a predesigned questionnaire including age, gender, occupation, residency and whether the drinking water was safe (purified) or not. Physical examination was done including weight status, BMI was calculated (BMI=wt. (kg)/ height (M) [2]. Provisional diagnosis of gallbladder disease was confirmed by examination, necessary laboratory investigations (Hematology, Biochemistry, and radiology). Cholecystectomy was done by using Laparoscopic cholecystectomy or open surgery. The presence of H.pylori in the mucosa of excised gallbladder was studied by using: 1) Rapid urease kit (HNAN C., LTD) 2) Histopathology & chemical analysis of associated gallstone Results: Seventy-eight patients undergoing cholecystectomy for symptomatic gallbladder disease, the gallbladder mucosa of 30 patients were tested positive for H. pylori with any one of the tests used in this study. The rapid urease test was sensitive 57.1% and specific 58.3 % of the cholecystectomies performed in our study. The mean age of studied patients was (34 ± 4 years). Females constitute 73.1% (57 out of the total 78 of patients). Of the studied cases; 26 patients (33.3%) were obese. Employee patients constitute 43 (55.1%) of patients, and 53 patients (67.9%) lived in urban areas. Purified water consumed by 61 (78.2%) of the studied population. Gallstones were detected in 56 (71.8%) of studied cases, acalculus chronically inflamed gallbladder was found in 20 (25.64%), and 2 cases (2.56%). were found to have gallbladder polyp. pylori infection was diagnosed in 30 (38.5%) of total cases (of the 56 calculus confirmed cases 48.2% were H.pylori positive, and of the 22 acalculus cases 13.6% H.pylori positive). Conclusion: Significant association is found between chronic calculus cholecystitis and H.pylori infection. While no significant association was found in correlation with acalculus cholecystitis and other gallbladder pathology. In regard to feasibility of using urease kit test, it is found that this test is sensitive 57.1%, and specific 58.3%, as such it is less accurate than histopathology study. Significant correlation was found between age, gender, weight status, and non-purified water source with H.pylori infection..

Download Full-text