scholarly journals Use of a Novel Enzyme Immunoassay Based on Detection of Circulating Antigen in Serum for Diagnosis of Helicobacter pylori Infection

2004 ◽  
Vol 11 (4) ◽  
pp. 775-779 ◽  
Author(s):  
Abdelfattah M. Attallah ◽  
Hisham Ismail ◽  
Gellan G. Ibrahim ◽  
Mohamed Abdel-Raouf ◽  
Ahmed M. El-Waseef ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Endoscopic Biopsy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Target Antigen ◽  
Serum Samples ◽  
Predictive Values ◽  
Biopsy Specimens ◽  
Significant Difference ◽  
Circulating Antigen ◽  
H Pylori

ABSTRACT Recently, noninvasive diagnostic tests for Helicobacter pylori infection have gained in significance. We have developed a sensitive and specific noninvasive immunoassay based on the detection of an H. pylori circulating antigen (HpCA) in sera from H. pylori-infected individuals. Monospecific antibody and Western blot analyses were used to demonstrate the presence of the target antigen in H. pylori cell lysate and serum samples. A novel enzyme-linked immunosorbent assay (ELISA) was developed for the detection of HpCA in serum. Endoscopic biopsy specimens from the gastric antra of 221 individuals (143 males and 78 females) with dyspeptic symptoms were evaluated for H. pylori infection, with culture used as a “gold standard” for diagnosis. The target H. pylori antigen was identified at 58 kDa. HpCA has been detected by ELISA with high degrees of sensitivity, specificity, and efficiency (>90%), and ELISA results show no significant difference (P > 0.05) from results of H. pylori culture of gastric biopsy specimens. The test's positive and negative predictive values were also high (95 and 86%, respectively). In conclusion, a sensitive and specific immunoassay was developed for the detection of HpCA in human serum. This test can be applied for noninvasive laboratory and field diagnoses of H. pylori infection.

scholarly journals Relationship of Anti-Lewis x and Anti-Lewis y Antibodies in Serum Samples from Gastric Cancer and Chronic Gastritis Patients toHelicobacter pylori-Mediated Autoimmunity

2001 ◽  
Vol 69 (8) ◽  
pp. 4774-4781 ◽  
Author(s):  
Michael A. Heneghan ◽  
Ciaran F. McCarthy ◽  
Daiva Janulaityte ◽  
Anthony P. Moran
Keyword(s):  
Gastric Cancer ◽  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Antibody Production ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Lewis Y ◽  
H Pylori ◽  
Negative Controls ◽  
Relationship Of

ABSTRACT Lewis (Le) antigens have been implicated in the pathogenesis of atrophic gastritis and gastric cancer in the setting ofHelicobacter pylori infection, and H. pylori-induced anti-Le antibodies have been described that cross-react with the gastric mucosa of both mice and humans. The aim of this study was to examine the presence of anti-Le antibodies in patients with H. pylori infection and gastric cancer and to examine the relationships between anti-Le antibody production, bacterial Le expression, gastric histopathology, and host Le erythrocyte phenotype. Anti-Le antibody production and H. pylori Le expression were determined by enzyme-linked immunosorbent assay, erythrocyte Le phenotype was examined by agglutination assays, and histology was scored blindly. Significant levels of anti-Lex antibody (P < 0.0001, T = 76.4, DF = 5) and anti-Ley antibody (P < 0.0001, T = 73.05, DF = 5) were found in the sera of patients with gastric cancer and other H. pylori-associated pathology compared with H. pylori-negative controls. Following incubation of patient sera with synthetic Le glycoconjugates, anti-Lex and -Ley autoantibody binding was abolished. The degree of the anti-Lex and -Leyantibody response was unrelated to the host Le phenotype but was significantly associated with the bacterial expression of Lex (r = 0.863,r 2 = 0.745, P < 0.0001) and Ley (r = 0.796,r 2 = 0.634, P < 0.0001), respectively. Collectively, these data suggest that anti-Le antibodies are present in most patients with H. pyloriinfection, including those with gastric cancer, that variability exists in the strength of the anti-Le response, and that this response is independent of the host Le phenotype but related to the bacterial Le phenotype.

scholarly journals Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

2013 ◽  
Vol 20 (10) ◽  
pp. 1569-1577 ◽  
Author(s):  
Abdelfattah M. Attallah ◽  
Faisal A. Bughdadi ◽  
Atef M. El-Shazly ◽  
Hisham Ismail
Keyword(s):  
Laboratory Diagnosis ◽  
Fasciola Gigantica ◽  
Antigen Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Target Antigen ◽  
Immunoperoxidase Staining ◽  
Serum Samples ◽  
Content Type ◽  
Human Fascioliasis ◽  
Circulating Antigen

ABSTRACTCurrently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test forFasciolainfection should be based on the detection of circulatingFasciolaantigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in aFasciola giganticaadult worm antigen preparation, excretory-secretory products, and sera fromF. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts ofF. giganticaadult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based onF. giganticacirculating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosedF. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC = 0.961,P< 0.0001) for discriminatingFasciola-infected and noninfected individuals. The developed assay showed high degrees of sensitivity, specificity, and efficiency (>93%), and a significant correlation (r= 0.715,P< 0.0001) between antigen level and parasite egg count was shown. In conclusion, a 27-kDaFasciolaantigen was identified in sera ofF. gigantica-infected individuals. A highly sensitive and specificFasciolaantigen detection assay, FgCA-27 ELISA, was developed for laboratory diagnosis of human fascioliasis.

scholarly journals Induction of various cytokines and development of severe mucosal inflammation by cagA gene positive Helicobacter pylori strains

Gut ◽  
1997 ◽  
Vol 41 (4) ◽  
pp. 442-451 ◽  
Author(s):  
Y Yamaoka ◽  
M Kita ◽  
T Kodama ◽  
N Sawai ◽  
K Kashima ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Expression Patterns ◽  
Mucosal Inflammation ◽  
Enzyme Linked Immunosorbent Assay ◽  
Biopsy Specimens ◽  
Tnf Α ◽  
Polymerase Chain ◽  
H Pylori ◽  
Factor Α ◽  
Necrosis Factor

Background—Helicobacter pyloristrains possessing the cagA gene are thought to induce interleukin 8 (IL-8) in gastric mucosa. However, it is still unclear whether a relation exists between the cagA gene and the expression patterns of cytokines other than IL-8.Aims—To investigate the relation between the cagA gene and the production of various cytokine proteins using an enzyme linked immunosorbent assay (ELISA).Patients and methods—In 184 patients, the cagA gene was detected by polymerase chain reaction (PCR), and levels of production of IL-1β, IL-6, IL-7, IL-8, IL-10, and tumour necrosis factor α (TNF-α) in antral biopsy specimens were measured by ELISA.Results—Mucosal levels of IL-1β, IL-6, IL-8, and TNF-α were significantly higher in H pyloripositive than in H pylori negative patients. Furthermore, the mucosal levels of IL-1β and IL-8 were significantly higher in specimens infected with cagApositive strains than in those infected with cagAnegative strains. In H pylori positive patients, the mucosal level of IL-8 was closely correlated with that of IL-1β (p<0.0001), and the mucosal level of IL-6 was closely correlated with that of TNF-α (p<0.0001).Conclusion—These findings suggest that the ability to induce cytokines differs among the strains;cagA+ strains induce various kinds of cytokines and may cause severe inflammation, whereascagA− strains induce IL-8 and IL-1β only weakly and may cause only mild inflammation. However, as most patients infected with the cagA+ strains have gastritis, these strains may not be equivalent to ulcerogenic strains.

scholarly journals Immunoblot Analysis of Humoral Immune Response to Helicobacter pylori in Children with and without Duodenal Ulcer

2000 ◽  
Vol 38 (5) ◽  
pp. 1777-1781 ◽  
Author(s):  
Gifone A. Rocha ◽  
Andreia M. R. Oliveira ◽  
Dulciene M. M. Queiroz ◽  
Anfrisina S. T. Carvalho ◽  
Ana M. M. F. Nogueira
Keyword(s):  
Immune Response ◽  
Duodenal Ulcer ◽  
Helicobacter Pylori ◽  
Humoral Immune Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Method ◽  
Predictive Values ◽  
Humoral Immune ◽  
Urease Test ◽  
H Pylori

Several studies have demonstrated that enzyme-linked immunosorbent assay is not a sensitive and specific method to diagnoseHelicobacter pylori infection in children, especially in the younger ones. Since serum immune response can also be determined by immunoblotting and it permits the detection of antibodies to virulence factors such as CagA and VacA, we evaluated the accuracy of a commercial immunoblotting test to diagnose H. pyloriinfection and to assess the humoral immune response to differentH. pylori antigens in 122 children who underwent upper gastrointestinal endoscopy. The presence of H. pylori was determined in antral biopsy specimens by culture, preformed urease test, and histological analysis. H. pylori was identified by microbiological and histopathological methods in 66 children (including all of the 21 who had duodenal ulcer). Antibodies toH. pylori were detected in 63 infected children and in 8 noninfected ones. The sensitivity, specificity, and positive and negative predictive values of the immunoblotting test were 95.5, 85.7, 88.7, and 94.1%, respectively. The number of immunoreactive bands increased with age (P = 0.003), and the bands of 35 kDa (P = 0.013); 89 kDa, the VacA antigen (P = 0.001); and 116 kDa, the CagA antigen (P = 0.00004) were more frequently observed in older children. The frequency of the bands of 89 kDa (P = 0.001) and 116 kDa (P = 0.03) was higher in children with duodenal ulcer than in H. pylori-positive children without the disease. In conclusion, the immunoblotting test appears to be useful for the diagnosis of H. pylori infection in children, even in the younger ones.

scholarly journals Detection of Anti-CagA Antibodies in Sera of Helicobacter pylori-Infected Patients Using an Immunochromatographic Test Strip

2019 ◽  
Vol 58 (3) ◽  
pp. 217-222
Author(s):  
Cebrail Karakus ◽  
Zeynep Ulupinar ◽  
Fahri Akbas ◽  
Duygu Yazici
Keyword(s):  
Helicobacter Pylori ◽  
Test Strip ◽  
Mucosal Damage ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Line ◽  
Immunochromatographic Test ◽  
Caga Gene ◽  
Serum Samples ◽  
Ulcer Disease ◽  
H Pylori

Abstract The cagA gene of Helicobacter pylori that encodes an immunodominant CagA protein provokes severe mucosal damage and acts as a risk factor for the development of peptic ulcer disease and gastric cancer. Our aim is to develop an immunochromatographic test strip (ICTS) using our previously developed recombinant CagA (rCagA) protein and anti-rCagA monoclonal antibody (Mab) for the detection of anti-CagA antibodies in sera of infected patients. The rCagA was firstly conjugated to gold nanoparticle and placed into the conjugate pad. A nonconjugated rCagA and anti-rCagA Mab (CK-02) were immobilized on the test line and control line, respectively. Biopsy and serum samples from 30 H. pylori-infected patients were used. The presence of cagA gene in biopsy samples was first detected by PCR (Polymerase Chain Reaction), and 22 patients were found positive while 8 were negative. When serum samples were tested by our developed ICTS, 21 were positive for anti-CagA antibodies while 9 were negative. The serum samples were also tested by a commercial ELISA (Enzyme Linked Immunosorbent Assay), and when compared to the ICTS a sensitivity of 95% and a specificity of 100% were obtained. The ICTS can be used for rapid detection of CagA-positive H. pylori infection instead of expensive, time consuming and laborious invasive approaches.

scholarly journals A Comparison of Accuracy between IMMULITE2000® and GENEDIA® for Helicobacter pylori Infection

2020 ◽  
Vol 20 (1) ◽  
pp. 54-62
Author(s):  
Seon Hee Lim ◽  
Nayoung Kim ◽  
Sung Eun Kim ◽  
Gwang Ho Baik ◽  
Ju Yup Lee ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Solid Phase ◽  
Pearson Correlation ◽  
Predictive Ability ◽  
Enzyme Linked Immunosorbent Assay ◽  
Operating Characteristics ◽  
Health Checkup ◽  
Serological Tests ◽  
Significant Difference ◽  
H Pylori

Background/Aims: In serological tests for <i>Helicobacter pylori</i> (<i>H. pylori</i>), an enzyme-linked immunosorbent assay (GENEDIA<sup>®</sup>) and a solid-phase, two-step chemiluminescent enzyme immunoassay (IMMULITE<sup>®</sup>), which are easy to perform, inexpensive, and widely available, are commonly used. However, local validation of the test performance of IMMULITE<sup>®</sup> is required. This study aimed to examine the performance of IMMULITE<sup>®</sup> in comparison with that of GENEDIA<sup>®</sup> in a Korean health checkup population.Materials and Methods: The sera of 300 subjects among those who underwent health checkup were analyzed using IMMULITE<sup>®</sup>, and results were compared with those of GENEDIA<sup>®</sup>. The two serological tests were compared for their ability to predict atrophic gastritis (AG) or intestinal metaplasia (IM) on endoscopy.Results: We found significant correlation (Pearson correlation coefficient=0.903, <i>P</i><0.0001) and an almost perfect agreement (Cohen’s Kappa coefficient=0.987, <i>P</i><0.0001) between the results of GENEDIA<sup>®</sup> and IMMULITE<sup>®</sup>. The area under the receiver operating characteristics curve (AUC) for AG using GENEDIA<sup>®</sup> and IMMULITE<sup>®</sup> were 0.590 and 0.604, respectively, and showed no statistically significant difference in predictive ability for AG (<i>Z</i>-statistics=-0.517, <i>P</i>=0.605). The AUC for IM by GENEDIA<sup>®</sup> and IMMULITE<sup>®</sup> were 0.578 and 0.593, respectively, with no statistically significant difference in predictive ability for IM between the two values (<i>Z</i>-statistics=-0.398, <i>P</i>=0.691).Conclusions: No statistically significant difference in diagnostic value for <i>H. pylori</i> infection was found between GENEDIA<sup>®</sup> and IMMULITE<sup>®</sup>.

scholarly journals Helicobacter pylori-Specific Immune Responses of Children: Implications for Future Vaccination Strategy

2002 ◽  
Vol 9 (5) ◽  
pp. 1126-1128 ◽  
Author(s):  
Günter Bode ◽  
Isolde Piechotowski ◽  
Dietrich Rothenbacher ◽  
Hermann Brenner
Keyword(s):  
Helicobacter Pylori ◽  
Confidence Interval ◽  
Immune Responses ◽  
Western Blot ◽  
Vaccination Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Significant Difference ◽  
Specific Igg ◽  
H Pylori

ABSTRACT We analyzed the specific anti-Helicobacter pylori immunoglobulin G (IgG) antibody profile for a sample of 824 asymptomatic schoolchildren in southern Germany (mean age, 10.7 ± 0.65 years) with an H. pylori-specific IgG enzyme-linked immunosorbent assay and Western blot analysis. The prevalence of infection was 19.8% (95% confidence interval, 17.1 to 22.7%). The immunoresponses were characterized predominantly by antibodies against low-molecular-mass antigens of 14 and 29 kDa, with a significant difference between children of German and Turkish nationalities (P = 0.0012 and P < 0.0001, respectively).

scholarly journals Chemokines in the gastric mucosa in Helicobacter pylori infection

Gut ◽  
1998 ◽  
Vol 42 (5) ◽  
pp. 609-617 ◽  
Author(s):  
Y Yamaoka ◽  
M Kita ◽  
T Kodama ◽  
N Sawai ◽  
T Tanahashi ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Expression Patterns ◽  
Enzyme Linked Immunosorbent Assay ◽  
Biopsy Specimens ◽  
Inflammatory Protein ◽  
Polymerase Chain ◽  
H Pylori ◽  
Macrophage Inflammatory Protein

Background—Although chemokines have been suggested to play an important role in Helicobacter pyloriassociated gastritis, few studies have investigated the role of chemokines other than interleukin 8 (IL-8) in gastric mucosa.Aims—To investigate the expression and production patterns of various chemokines using gastric biopsy specimens.Methods—In 192 patients, expression patterns of C-X-C chemokines (IL-8 and growth regulated α (GROα)) and C-C chemokines (regulated on activation, normal T cell expressed and presumably secreted (RANTES), monocyte chemotactic and activating factor (MCAF), macrophage inflammatory protein 1α (MIP-1α), and MIP-1β) were examined using reverse transcription polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA).cagA gene was identified using PCR.Results—H pylori infection was associated with increased rates of expression of mRNA for IL-8, GROα, RANTES, and MIP-1α and with increased levels of mucosal IL-8 and GROα. IL-8 and GROα levels correlated with the density of H pylori in both the antrum and corpus. The levels of these chemokines correlated with cellular infiltration in the antrum but not the corpus. cagA gene positive H pyloriinfection was associated with increased rates of expression of mRNA for IL-8 and GROα and with increased levels of these chemokines.Conclusion—H pylori infection is associated with increased expression rates and production of C-X-C chemokines (IL-8 and GROα), but not with increased production of C-C chemokines. Although H pylori infection is associated with increased C-X-C chemokines in the antrum and corpus, there is a difference in the inflammatory response between these two areas of the stomach.

scholarly journals The burden of Helicobacter pylori infection in England and Wales

2002 ◽  
Vol 128 (3) ◽  
pp. 411-417 ◽  
Author(s):  
A. J. VYSE ◽  
N. J. GAY ◽  
L. M. HESKETH ◽  
N. J. ANDREWS ◽  
B. MARSHALL ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
High Reactivity ◽  
Antimicrobial Treatment ◽  
Active Infection ◽  
Serum Samples ◽  
England And Wales ◽  
Significant Difference ◽  
Specific Igg ◽  
H Pylori ◽  
Serum Elisa

The prevalence of active infection with Helicobacter pylori in the general population of England and Wales was estimated using high reactivity for specific IgG in serum ELISA as a marker. A total of 10118 anonymized residues of serum samples collected in 1986 and 1996 from persons aged 1–84 years were used. Estimated prevalence of active infection varied by region and was highest in London. Prevalence was related to decade of birth and increased from 4·3% in those born during the 1980s to 30% in those born before 1940. An estimated total of 7·5 million people living in England and Wales have an active infection and analysis by decade of birth showed no significant difference between samples collected in 1986 and 1996. These data suggest H. pylori infection is becoming less common, is acquired at an early age and is unlikely to be resolved unless suitable antimicrobial treatment is sought.

scholarly journals Helicobacter Pylori Infection in Endoscopic Biopsy Specimens of Gastric Cancer: A Preliminary Evaluation in a High Risk Population of Kashmir Valley

1997 ◽  
Vol 4 (1) ◽  
pp. 35-42 ◽  
Author(s):  
G. M. Malik ◽  
S. Kadla ◽  
M. Mubarik ◽  
T. Hussain ◽  
G. Jeelani ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Helicobacter Pylori ◽  
Dietary Habits ◽  
Endoscopic Biopsy ◽  
High Risk Population ◽  
Preliminary Evaluation ◽  
Kashmir Valley ◽  
Biopsy Specimens ◽  
Kashmiri Population ◽  
H Pylori

Objective: The aim of the present study was to assess the prevalence of Helicobacter pylori (H. pylori) infection in Kashmiri patients with gastric cancer and to compare this with a matched control population.Methods: Fifty patients with gastric cancer and thirty age/sex matched controls were included in the study. All the subjects were hailing from Kashmir Valley. For detection of H. pylori, biopsy specimens were used both from cases and controls.Results: An insignificant association was shown between H. pylori and both intestinal and diffuse type of gastric cancer.Conclusions: The data provides support against the significant association between H. pylori and gastric cancer in this part of world, a place where the age standardized incidence of gastric cancer is alarmingly high. We conclude that other factors like personal and special dietary habits of Kashmiri population may be more important for the development of gastric cancer.

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