Journals / Mikrobiyoloji Bülteni / 2010 / Cilt: 44 - Sayı: 1

Evaluation of Western Blot method for the detection of antibodies to Helicobacter pylori antigens in patients with gastric carcinoma and cases with epigastric complaints

Gastrik karsinomalı hastalar ile epigastrik yakınmaları olan olgularda Helicobacterpylori antijenlerine karşı antikor varlığının saptanmasında VVestern Blot yönteminin değerlendirilmesi

Pages
21–28
DOI
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AI Summary & Key Takeaways

AI Insight
⚡ TL;DR(Too Long; Didn't Read)

These results suggested that this test should be assessed again by the manufacturer for its detection power directed towards specific H.pylori antibodies, especially for Vac-A.

📌 3 Key Takeaways

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Objective & Hypothesis:Helicobacter pylori proteins CagA (cytotoxin-associated gene A) and VacA (vacuolating cytotoxin A) are among the virulence factors of this species.
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Key Findings & Data:A total of 99 patients with gastric cancer (94 adenocarcinoma, 2 adenosqu-amous cell carcinoma, 3 non-Hodgkin lymphoma) and 150 control cases with epigastric complaints such as nausea, vomiting, diarrhea, gastroesophageal reflux and abdominal pain, were included to the study.
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Conclusion & Impact:These results suggested that this test should be assessed again by the manufacturer for its detection power directed towards specific H.pylori antibodies, especially for Vac-A.
🔬Design:Research Article
📍Scope:Comprehensive Sample
⚙️Variables:Domain Parameters

Abstract

Helicobacter pylori proteins CagA (cytotoxin-associated gene A) and VacA (vacuolating cytotoxin A) are among the virulence factors of this species. CagA gene carrying H.pylori strains are particularly associated with gastric adenocarsinoma. This study was conducted to evaluate Western Blot (WB) method to determine specific H.pylori antibodies in a group of patients with gastric cancer and in a control group with no malignancy. A total of 99 patients with gastric cancer (94 adenocarcinoma, 2 adenosqu-amous cell carcinoma, 3 non-Hodgkin lymphoma) and 150 control cases with epigastric complaints such as nausea, vomiting, diarrhea, gastroesophageal reflux and abdominal pain, were included to the study. H.pylori IgG-ELISA was positive in all study (mean age: 56.7 ± 1.2 years, 62 male) and control (mean age: 24.2 ± 1.3 years, 64 male) patients. Specific antibodies against CagA, VacA, OMP (outer membrane protein)-67, urease-A, urease-B, HSP (heat shock protein) and flagellin antigens determined by a commercial WB-based kit (RIDA Blot Helicobacter, R-Biopharm GmbH, Germany). Interestingly, no anti-VacA positivity was detected in none of the patient and control groups. The positivity rates for H.pylori CagA, OMP-67, urease A, urease-B, flagellin and HSP specific antibodies were as 78%, 54%, 37%, 60%, 53% and 82% in the gastric cancer group and 85%, 71%, 55%, 43%, 61% and 75% in the control group, respectively. There was no statistically significant difference (p> 0.05) between gastric carcinoma and control groups in terms of CagA, HSP and flagellin antibodies (p> 0.05). On the other hand, a statistically significant difference was found between the 2 groups in terms of urease-A, urease-B and OMP-67 (p< 0.01). These results suggested that this test should be assessed again by the manufacturer for its detection power directed towards specific H.pylori antibodies, especially for Vac-A. Further molecular and clinical studies are necessary to determine the factors that affect H.pylori virulence and disease prognosis.

Özet

Helicobacter pylori VacA (vacuolating cytotoxin A) ve CagA (cytotoxin associated gene A) proteinleri, bakterinin virülans faktörleri arasında yer almaktadır. Özellikle CagA geni taşıyan H.pylori suşlarının gastrik adenokarsinom gelişiminde potansiyel risk faktörü olduğu belirtilmektedir. Bu çalışmada, mide kanseri tanısı almış hastalarda ve malignansisi olmayan kontrol grubunda çeşitli H.pylori antijenlerine karşı özgül antikorların saptanmasında Western Blot (WB) yönteminin değerlendirilmesi amaçlanmıştır. Çalışmaya, kanserli 99 hasta (94 adenokarsinom, 2 adenoskuamöz hücreli karsinom, 3 non-Hodgkin lenfo-ma) ile kontrol olarak bulantı, kusma, ishal, regürjitasyon ve karın ağrısı gibi epigastrik şikayetleri olan 150 olgu alınmıştır. Hasta (yaş ortalaması: 56.7 ±1.2 yıl; 62'si erkek) ve kontrol (yaş ortalaması: 24.2 ± 1.3 yıl; 64'ü erkek) gruplarındaki tüm olgularda ELISA yöntemiyle H.pylori IgG pozitifliği mevcuttur. H.pylori CagA, VacA, OMP-67 (outer membrane protein), üreaz A, üreaz B, flajellin ve HSP (heat shock protein) antijenlerine karşı antikor varlığı, ticari bir test (RIDA Blot Helicobacter; R-Biopharm GmbH, Almanya) kullanılarak araştırılmıştır. Çalışmamızda, ilginç olarak hasta ve kontrol bireylerin hiçbirisinde an-ti-VacA pozitifliği saptanmamış; CagA, OMP, üreaz A, üreaz B, flajellin ve HSP'ye özgül antikor pozitiflik oranları hasta grubunda sırasıyla; %78, %54, %37, %60, %53 ve %82; kontrollerde ise sırasıyla %85,%71, %55, %43, %61 ve %75 olarak tespit edilmiştir. Hasta ve kontrol grupları arasında CagA, HSP ve flajellin antikorlarının pozitiflik oranları istatistiksel olarak anlamlı bir fark göstermezken (p> 0.05); anti-OMP67, anti-üreaz A ve anti-üreaz B pozitiflik oranları arasında anlamlı bir fark saptanmıştır (p< 0.01). Sonuç olarak, H.pylori antijenlerine karşı oluşan özgül antikorların saptanması ve ayırt edilmesinde WB temelli bu testin, özellikle vacA antikorlarını saptama açısından üretici firma tarafından bir kez daha değerlendirilmesinin gerekli olduğu kanaatine varılmış ve H.pylori'nm virülansını ve enfeksiyonun prognozunu belirlemede önemi olan faktörlerin aydınlatılmasında daha ileri moleküler ve klinik çalışmalara gereksinim olduğu düşünülmüştür.

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Cite This Article

GÜDÜCÜOĞLU, H., BERKTAŞ, M., BOZKURT, H., ÖZER, T. T., BULUT, G., ÖZTÜRK, Ö., İLHAN, M. (2010). Evaluation of Western Blot method for the detection of antibodies to Helicobacter pylori antigens in patients with gastric carcinoma and cases with epigastric complaints. Mikrobiyoloji Bülteni, Cilt: 44 - Sayı: 1, 21–28. https://ajindex.com/articles/298810

References (13)

  1. Brown LM. Helicobacter pylori: epidemiology and routes of transmission. Epidemiol Rev 2000
  2. Sugiyama T. Development of gastric cancer associated with Helicobacter pylori infection. Cancer Chemot-her Pharmacol 2004
  3. Sezikli M, Guliter S, Apan TZ, Aksoy A, Keles H, Ozkurt ZN. Frequencies of serum antibodies to Helicobacter pylori CagA and VacA in a Turkish population with various gastroduodenal diseases. Int J Clin Pract 2006
  4. Akhter Y, Ahmed I, Devi SM, Ahmed N. The co-evolved Helicobacter pylori and gastric cancer: trinity of bacterial virulence, host susceptibility and lifestyle. Infect Agent Cancer 2007
  5. Bülent K, Murat A, Esin A, et al. Association of CagA and VacA presence with ulcer and non-ulcer dyspepsia in a Turkish population. World J Gastroenterol 2003
  6. Ece F, F Hatabay N, Erdal N, Gedik C, Güney C, Aksoy F. Does Helicobacter pylori infection play a role in lung cancer? Respir Med 2005
  7. laquinto G, Todisco A, Giardullo N, et al. Antibody response to Helicobacter pylori CagA and heat-shock proteins in determining the risk of gastric cancer development. Dig Liver Dis 2000
  8. Mitchell HM, Hazell SL, Li YY, Hu PJ. Serological response to specific Helicobacter pylori antigens: antibody against CagA antigen is not predictive of gastric cancer in a developing country. Am J Gastroenterol 1996
  9. Lamarque D, Gilbert T, Roudot-Thoraval F, Deforges L, Chaumette MT, Delchier JC. Seroprevalence of eight Helicobacter pylori antigens among 182 patients with peptic ulcer, MALT gastric lymphoma or non-ulcer dyspepsia. Higher rate of seroreactivity against CagA and 35-kDa antigens in patients with peptic ulcer originating from Europe and Africa. Eur J Gastroenterol Hepatol 1999
  10. Elitsur Y, Neace C, Werthammer MC, Triest WE. Prevalence of CagA, VacA antibodies in symptomatic and asymptomatic children with Helicobacter pylori infection. Helicobacter 1999
  11. Covacci A, Censini S, Bugnoli M, et al. Molecular characterization of the 128-kDa immunodominant antigen of Helicobacter pylori associated with cytotoxicity and duodenal ulcer. Proc Natl Acad Sci USA 1993
  12. Schumann C, Triantafilou K, Rasche FM, et al. Serum antibody positivity for distinct Helicobacter pylori antigens in benign and malignant gastroduodenal disease. Int j Med Microbiol 2006
  13. Wong BC, Lam SK, Ching CK, et al. Seroprevalence of cytotoxin-associa