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

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.

1999 ◽  
Vol 6 (4) ◽  
pp. 489-493 ◽  
Author(s):  
Guillermo I. Perez-Perez ◽  
Richard M. Peek ◽  
John C. Atherton ◽  
Martin J. Blaser ◽  
Timothy L. Cover

ABSTRACT Several different families of vacuolating toxin (vacA) alleles are present in Helicobacter pylori, and they encode products with differing functional activities. H. pyloristrains containing certain types of vacA alleles have been associated with an increased risk for peptic ulcer disease. In this study, we tested serum samples and gastric juice from 19 H. pylori-negative and 39 H. pylori-positive patients for enzyme-linked immunosorbent assay reactivity with two different types of VacA antigens (types s1/m1 and s2/m2), which were purified from H. pylori 60190 and 86-338, respectively. Both antigens were recognized better by serum immunoglobulin G (IgG) fromH. pylori-positive persons than by serum IgG from H. pylori-negative persons (P < 0.01). The s1/m1 VacA antigen was better recognized by sera from patients carryingvacA type s1/m1 strains than by sera from patients carryingvacA type s2/m2 or s1/m2 strains (P < 0.01). Conversely, the s2/m2 VacA antigen was better recognized by sera from patients carrying type s2/m2 or s1/m2 strains (P= 0.03). Serum IgG anti-VacA antibodies were present more frequently in patients carrying type s1/m1 strains than in other H. pylori-positive patients (P = 0.0002). In addition, the highest levels of IgA anti-VacA antibodies were detected in the gastric juice of patients carrying type s1/m1 strains. These data indicate that different VacA isoforms have distinct antigenic properties and that multiple forms of VacA elicit antibody responses inH. pylori-positive humans.


2001 ◽  
Vol 69 (8) ◽  
pp. 4774-4781 ◽  
Author(s):  
Michael A. Heneghan ◽  
Ciaran F. McCarthy ◽  
Daiva Janulaityte ◽  
Anthony P. Moran

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.


Gut ◽  
1997 ◽  
Vol 41 (2) ◽  
pp. 169-176 ◽  
Author(s):  
S Rosenstock ◽  
L Kay ◽  
C Rosenstock ◽  
L P Andersen ◽  
O Bonnevie ◽  
...  

Background—Helicobacter pylori is a human pathogen that colonises the gastric mucosa and causes permanent gastric inflammation.Aims—To assess the symptoms of H pylori infection in an adult unselected population.Subjects—A random sample of 3589 adult Danes who were examined in 1982 and 1987 (n=2987).Methods—Abdominal symptoms within the preceding year were recorded at both attendances. Circulating IgG antibodies against H pylori in serum samples drawn in 1982 were measured by using in-house indirect enzyme linked immunosorbent assays (ELISA).Results—People with increased levels of IgG antibodies to H pylori were more likely than uninfected individuals to report heartburn (odds ratio (OR) = 1.26, 95% confidence interval (CI) 1.03–1.54) and abdominal pain characterised by daily length (OR = 1.33, 95% CI 0.92–1.91), nocturnal occurrence (OR = 1.62, 95% CI 1.19–2.19), spring aggravation (OR = 1.68, 95% CI 0.70–4.05), and no relation to meals (OR = 0.62, 95% CI 0.43–0.91) or stress (OR = 0.69, 95% CI 0.50–0.95). The inclusion of people with increased levels of IgG antibodies to H pylori, but without upper dyspepsia, at study entry significantly increased the likelihood of reporting upper dyspepsia at follow up (OR = 1.71, 95% CI 1.24–2.36). People with epigastric pain and increased levels of IgM antibodies to H pylori only indicative of acute H pylori infection were more likely to report nocturnal pain, heartburn, nausea, and vomiting.Conclusions—H pylori infection may precede the development of dyspepsia and is associated with a variety of gastrointestinal symptoms in people with no history of peptic ulcer disease.


2004 ◽  
Vol 11 (4) ◽  
pp. 775-779 ◽  
Author(s):  
Abdelfattah M. Attallah ◽  
Hisham Ismail ◽  
Gellan G. Ibrahim ◽  
Mohamed Abdel-Raouf ◽  
Ahmed M. El-Waseef ◽  
...  

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.


2015 ◽  
Vol 9 (10) ◽  
pp. 1108-1116 ◽  
Author(s):  
Ivy Bastos Ramis ◽  
Júlia Silveira Vianna ◽  
Priscila Cristina Bartolomeu Halicki ◽  
Caroline Lara ◽  
Thássia Fernanda Tadiotto ◽  
...  

Introduction: Helicobacter pylori infection is associated with gastritis, peptic ulcer disease and gastric carcinoma. The severity of damage is determined by the interplay between environmental/behavioral factors, bacterial pathogenicity genes and host genetic polymorphisms that can influence the secretion levels of inflammatory cytokines. Accordingly, this study aimed to identify polymorphisms in the IL-1B and IL-1RN genes and their associations with H. pylori infection, cagA gene of H. pylori, and gastroduodenal diseases. Methodology: Gastric biopsy samples from 151 patients infected with H. pylori and 76 uninfected individuals were analyzed. H. pylori infection was diagnosed by histology and PCR. Polymorphisms at positions -511, -31 and +3954 of the IL-1B gene were detected by PCR-RFLP, and an analysis of the VNTR polymorphism of the IL-1RN gene was performed by PCR. Results: It was observed that the presence of the T/T genotype at position -511 and the C/C genotype at position -31 were associated with H. pylori infection and with an increased risk of gastritis in H. pylori-positive patients. Additionally, strains from patients H. pylori-positive carrying the cagA gene was significantly related with the T/T genotype at position -511 of IL-1B.  No association of polymorphisms at position +3954 of IL-1B and in the IL-1RN with H. pylori infection and with risk of severe gastric diseases was found. Conclusions: We demonstrated that polymorphisms in the promoter region of the IL-1B gene (at positions -511 and -31) are associated with an enhanced risk of H. pylori infection as well as gastritis in H. pylori-positive patients.


2020 ◽  
Vol 14 (1) ◽  
pp. 36-40
Author(s):  
Fahmida Rahman ◽  
Khandaker Shadia ◽  
Salma Khatun ◽  
Mafruha Mahmud ◽  
Indrajit Kumar Dutta ◽  
...  

Background: CagA IgG antibody in sera might indicate presence of virulent Helicobacter pylori in patients with peptic ulcer disease. Present study was performed to find out the prevalence of CagA IgG antibody in patients with peptic ulcer/erosion. Methods: Any case that had peptic ulcer/erosion, plus positive for rapid urease test (RUT) or H. pylori stool antigen (HpSAg) or serum anti-H. pylori IgG/IgA were included in the study and named as H. pylori positive case. H. pylori positive cases were tested for CagA IgG antibody. Anti-H. pylori IgG, IgA and CagA IgG antibodies were determined by enzyme-linked immunosorbent assay (ELISA) and stool antigen by rapid immunochromatographic test (ICT). Urease production in biopsy sample was detected by RUT. Results: Total 86 H. pylori positive patients were included in the study. Out of 86 patients, CagA IgG was positive in 34 (39.5%; 95% CI: 0.30,0.50) cases. CagA seropositivity rate in ulcer and erosion cases were 58.8% (95% CI: 0.36,0.78) and 34.8% (95% CI: 0.25,0.47) respectively. H. pylori stool antigen and IgA antibodies were positive in all (100%) CagA antibody positive ulcer cases while the rates were significantly less among the CagA antibody negative cases (42.8% and 28.6%; p<0.05). However, in CagA antibody positive erosion cases, the rates were not significantly different from CagA antibody negative cases. Conclusion: The study has demonstrated that the CagA positive strain is less prevalent in erosion than ulcer cases. Ibrahim Med. Coll. J. 2020; 14(1): 36-40


1996 ◽  
Vol 82 (1) ◽  
pp. 40-44
Author(s):  
Chew-Wun Wu ◽  
Tzee-Chung Wu ◽  
Yun-Ray Chang ◽  
Shyh-Haw Tsay ◽  
Shih-Jiun Yin ◽  
...  

We examined the biologic tumor behavior in Helicobacter pylori-seropositive patients with gastric adenocarcinoma. A total of 214 consecutive patients with pathologically confirmed adenocarcinoma of the stomach who underwent gastric resection were studied. The stored serum samples were tested for serum antibody to H. pylori by using a highly sensitive and specific IgG enzyme-linked immunosorbent assay. The difference in H. pylori-seropositive and seronegative patients with gastric adenocarcinoma was evaluated in terms of various clinicopathologic parameters. A multivariate logistic regression analysis was used to adjust for potential confounding variables. Antibodies to H. pylori were detected in 65.9% of patients with gastric adenocarcinoma. H. pylori-seropositive patients were younger than seronegative patients and had infiltrative tumor according to Ming's criteria. When adjusted for age, infiltrative tumor come out stronger. These findings suggest that H. pylori infection may be related to infiltrative type gastric adenocarcinoma; further study is necessary.


1998 ◽  
Vol 44 (2) ◽  
pp. 293-298 ◽  
Author(s):  
Wayne H Schrier ◽  
Ronald J Schoengold ◽  
Josefina T Baker ◽  
Joyce L Norell ◽  
Corey L Jaseph ◽  
...  

Abstract We describe a solid-phase immunochromatographic serologic test, FlexSure® HP, to detect IgG antibodies against Helicobacter pylori. H. pylori colonize the stomach and proximal duodenum, cause ulcer disease and mucosa-associated lymphoid tissue lymphoma, and have a role in the development of other disorders, including gastric adenocarcinoma. FlexSure HP consists of a test strip, conjugate pad, and absorbent pad, in a novel reverse-flow chromatography format. In these studies, FlexSure HP was demonstrated to be specific for IgG antibodies against H. pylori. The reactive cutoff of the test was consistent with [13C]urea breath test and commercially available ELISAs. FlexSure HP had 94% sensitivity, 88% specificity, and 91% accuracy relative to [13C]urea breath test; and 95% sensitivity, 94% specificity, and 95% overall agreement relative to high-molecular-mass cell-associated protein enzyme immunoassay (HM-CAP EIA). FlexSure HP is a simple-to-perform, visually read test requiring no specialized training, equipment, or instrumentation, and yields rapid, accurate, qualitative results.


2013 ◽  
Vol 20 (11) ◽  
pp. 1703-1710 ◽  
Author(s):  
Luca Formichella ◽  
Laura Romberg ◽  
Christian Bolz ◽  
Michael Vieth ◽  
Michael Geppert ◽  
...  

ABSTRACTHelicobacter pyloricolonizes half of the world's population, and infection can lead to ulcers, gastric cancer, and mucosa-associated lymphoid tissue (MALT) lymphoma. Serology is the only test applicable for large-scale, population-based screening, but current tests are hampered by a lack of sensitivity and/or specificity. Also, no serologic test allows the differentiation of type I and type II strains, which is important for predicting the clinical outcome.H. pylorivirulence factors have been associated with disease, but direct assessment of virulence factors requires invasive methods to obtain gastric biopsy specimens. Our work aimed at the development of a highly sensitive and specific, noninvasive serologic test to detect immune responses to importantH. pylorivirulence factors. This line immunoassay system (recomLine) is based on recombinant proteins. For this assay, six highly immunogenic virulence factors (CagA, VacA, GroEL, gGT, HcpC, and UreA) were expressed inEscherichia coli, purified, and immobilized to nitrocellulose membranes to detect serological immune responses in patient's sera. For the validation of the line assay, a cohort of 500 patients was screened, of which 290 (58.0%) wereH. pylorinegative and 210 (42.0%) were positive by histology. The assay showed sensitivity and specificity of 97.6% and 96.2%, respectively, compared to histology. In direct comparison to lysate blotting and enzyme-linked immunosorbent assay (ELISA), therecomLine assay had increased discriminatory power. For the assessment of individual risk for gastrointestinal disease, the test must be validated in a larger and defined patient cohort. Taking the data together, therecomLine assay provides a valuable tool for the diagnosis ofH. pyloriinfection.


2006 ◽  
Vol 74 (7) ◽  
pp. 4064-4074 ◽  
Author(s):  
Mónica Oleastro ◽  
Lurdes Monteiro ◽  
Philippe Lehours ◽  
Francis Mégraud ◽  
Armelle Ménard

ABSTRACT Peptic ulcer disease (PUD) occurs after a long-term Helicobacter pylori infection. However, the disease can develop earlier, and rare cases have been observed in children, suggesting that these H. pylori strains may be more virulent. We used suppressive subtractive hybridization for comparative genomics between H. pylori strains isolated from a 5-year-old child with duodenal ulcer and from a sex- and age-matched child with gastritis only. The prevalence of the 30 tester-specific subtracted sequences was determined on a collection of H. pylori strains from children (15 ulcers and 30 gastritis) and from adults (46 ulcers and 44 gastritis). Two of these sequences, jhp0562 (80.0% versus 33.3%, P = 0.008) and jhp0870 (80.0% versus 36.7%, P = 0.015), were highly associated with PUD in children and a third sequence, jhp0828, was less associated (40.0% versus 10.0%, P = 0.048). Among adult strains, none of the 30 sequences was associated with PUD. However, both jhp0562 and jhp0870 were less prevalent in adenocarcinoma strains than in PUD strains from children and adults, the difference being statistically significant for jhp0870. In conclusion, two H. pylori genes were identified as being strongly associated with PUD in children, and their putative roles as an outer membrane protein for jhp0870 and in lipopolysaccharide biosynthesis for jhp0562, suggest that they may be novel virulence factors of H. pylori.


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