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Use of serum immunoglobulins G and A for detection of Helicobacter pylori infection in dyspeptic patients by enzyme immunosorbent assay Babay HA, Al Mofleh IA, Al Akwaa AM, Al Humayed SM, Al Habbal MT - Saudi J Gastroenterol
Saudi Journal of Gastroenterology
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ARTICLES Table of Contents   
Year : 2000  |  Volume : 6  |  Issue : 1  |  Page : 33-36
Use of serum immunoglobulins G and A for detection of Helicobacter pylori infection in dyspeptic patients by enzyme immunosorbent assay


1 Department of Pathology/Microbiology, King Khalid University Hospital, Riyadh, Saudi Arabia
2 Department of Medicine/Gastroenterology Div., King Khalid University Hospital, Riyadh, Saudi Arabia

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Date of Submission 15-Oct-1998
Date of Acceptance 24-Aug-1999
 

   Abstract  

Since the discovery of Helicobacter pylori (H. pylori), several invasive and non invasive tests have become available. The aim of this study was to test the performance of immunoglobulins IgG and IgA by using an enzyme linked immunosorbent assay (ELISA) test (In vitro diagnostika GmbH, Germany) for the diagnosis of H. pylori among dyspeptic patients. Blood samples from 152 dyspeptic patients and 51 asymptomatic controls were analyzed in a case control study. IgG and IgA were positive in 33.5% and 41.1% respectively compared to 13.8% for both IgG and IgA in controls (P=0.002). We support the future use of serology as a non invasive, and rapid test for the diagnosis of H. pylori infection among dyspeptic patients in areas with low prevalence. Endoscopy remains the method of choice for elderly dyspeptic patients and for those with possible gastric or duodenal pathology.

Keywords: Helicobacter pylori, Serology, Immunoglobulines IgG, IgA.

How to cite this article:
Babay HA, Al Mofleh IA, Al Akwaa AM, Al Humayed SM, Al Habbal MT. Use of serum immunoglobulins G and A for detection of Helicobacter pylori infection in dyspeptic patients by enzyme immunosorbent assay. Saudi J Gastroenterol 2000;6:33-6

How to cite this URL:
Babay HA, Al Mofleh IA, Al Akwaa AM, Al Humayed SM, Al Habbal MT. Use of serum immunoglobulins G and A for detection of Helicobacter pylori infection in dyspeptic patients by enzyme immunosorbent assay. Saudi J Gastroenterol [serial online] 2000 [cited 2014 Mar 4];6:33-6. Available from: http://www.saudijgastro.com/text.asp?2000/6/1/33/33502


In 1983 Warren and Marshall introduced  Helicobacter pylori Scientific Name Search ori) as an important pathogen in human type - B gastritis and peptic ulcer disease[1],[2]. Since then, a major break through has come in the understanding of the pathogenesis of H. pylori infection. The interest in H. pylori has extended further after its being implicated in the pathogenesis of gastric adenocarcinoma and gastric lymphoma[3],[4]. Several methods have been used to detect H. pylori including isotope based urea breath test. For reasons that include the invasiveness of the endoscopic procedure, and high cost of urea breath test, the interest has been focused on the less invasive and less expensive serological techniques for the diagnosis of H. pylori infection.

A number of reports have emerged regarding the association between H. pylori and chronic gastritis, peptic ulcer and other upper gastrointestinal diseases among Saudi population[5],[6],[7]. Different methods of diagnosis were used including serology in few reports[8],[9]. The aim of the current study is to examine the effectiveness of serological tests using ELISA in the diagnosis of H. pylori infection in adult patients with dyspeptic symptoms in a case­control study.


   Subjects and Methods   Top


This prospective study was conducted between May and June 1997 at the Endoscopy Unit of KKUH, Riyadh, Saudi Arabia on 152 adult Saudi patients with dyspeptic symptoms. Their age ranged between 18 and 85 years. Fifty-one healthy male Saudi blood donors, aged between 35-59 years, served as a control group. Patients and controls were interviewed by a structured questionnaire regarding demographic details, past history and family history of peptic ulcer disease, any current illnesses, and dyspeptic symptoms. Patients who received antibiotics or bismuth in the preceeding two months, those on H2 receptor antagonists in the preceeding two weeks and those who had partial or complete gastrectomy were excluded from the study.

Blood samples were collected by venipuncture from patients and controls. Sera were separated and stored at -20 °C until assayed. Anti- H. pylori IgG and IgA antibodies were measured in all samples by an enzyme immunosorbentassay (ELISA) test using specific antigen (In vitro diagnostika GmbH, Germany). The assay was performed according to the manufacturer's instructions. For IgG, reading in the range 155-230 U/ml were considered positive, (>230 U/ml as high positive) and <135 U/ml as negative. For IgA, readings in the range 50-75 U/ml were considered positive, (>75 U/ml as high positive) and <40 U/ml as negative.


   Statistical Analysis   Top


Data were analysed by Chi-square. P value of <0.05 was considered significant.


   Results   Top


The median age of the patients and controls was 36 and 41 years, respectively. In the majority, dyspeptic pain was of motility or ulcer type of pain being 45.6% and 38%, respectively. There was no significant association between the symptoms and other patients characteristics. None of the control group had dyspeptic symptoms.

[Table - 1] presents the serologic findings among patients and control. For reason of easy interpretation, the positive and high positive results for both IgG and IgA were included together as positive. Patients with ulcer like symptoms had positive IgA (P=<0.05). Patients had significantly higher rate of positive IgA and IgG serology (P<0.002) compared to controls.


   Discussion   Top


Several investigators have documented the presence of IgG and IgA antibodies in individuals with H. pylori compared to those with normal gastric mucosa[10],[11],[12],[13]A variety of immunological techniques have been used for the rapid detection of H. pylori infection using monoclonal antibodies, like immunofluorescence and latex agglutination[8],[13]. For the diagnosis of H. pylori infection, ELISA assay has been accurate, non-invasive and easy to perform. Few studies have compared a large number of kits and some may have resulted in an artificially low specifities since the positive and negative predictive values of a test depend on the prevalence of the condition in the sample population[14]

The seropositivity of H. pylori IgG antibodies in our study was 33.5% compared to 13.8% among the control group. Wilcox, et al. 1996 reported a 35% prevalence[14]. In another study, utilizing high molecular weight cell associated protein (HM-CAP) H. pylori Immunoassay (EPI-enteric products, incorporated) found H. pylori IgG and IgA in 55% of 20 children with chronic epigastric pain [15] Mohamed et al. reported similar figures[9]. In contrast, a poor correlation between H. pylori infection and serology has been found when Serion H. pylori immunotab(Serion, Wurzberg, West Germany) and Pyloragen H. pylori (Hypcor Biomedical Inc. Irvine, CA) test kits were used[16] In another study screening asymptomatic Saudi population, rapid latex-agglutination test (Pylori-set, Orion Diagnostica, Epsoo, Finland) was of limited sensitivity. The author recommended ELISA instead of latex-agglutination test [8].

Other investigators reported serology to be sensitive, specific and non-invasive and have recommended it for screening dyspeptic patients under the age of 50 years[17],[18]. In addition to its accuracy for peptic ulcer patients, serology has resulted in reduction of endoscopy workload and cost[14],[19]. The low positive rates obtained by serology in our study could be due to the extensive awareness of population and possible earlier eradication. Spontaneous eradication and other factors have suggested by Andersen et al. They have correlated symptoms in seronegative patients with factors other than ulcers such as tobacco smoking, ingestion of nonsteroidal anti inflammatory drugs, or heriditary predisposition[20]. If eradication treatment is beneficial in non-ulcer dyspepsia, a policy of eradication in all patients with positive serology could be justified and thereby avoid the need for endoscopy[21]

The diagnostic efficacy of the ELISA test is highly dependent on the H. pylori antigen used. Our ELISA test used a specific low molecular weight H. pylori antigen with no nonspecific cross reactions.

In the current study, ELISA test showed that 33.5% of patients were IgG positive which might explain prior exposure to the bacteria. Those with positive IgA (41.1%) had recent infection or reinfection with H. pylori.

Detection of serum IgG antibodies has been sensitive (85-95%) and specific for detecting H. pylori carrier status[22]. The detection of H. pylori specific antibodies is consistent with a prolonged chronic mucosal inflammation with predominating both IgA and IgG[23]. Jaskowski et al study have shown that IgA may appear earlier than IgG antibodies in patients reinfected after unsuccessful treatment with antibiotics[24]. Clinical utility of IgA has shown that in the presence of IgG, IgA correlates with an acute infection in 95% and 74% of duodenal and gastric ulcer, respectively[25]. Yamamoto et al reported IgA specifity of 100% for H. pylori infection compared with CLO test, culture and histology[26]. Jaskowski et al. recommended that positive IgA and negative IgG in symptomatic patients may be of significant clinical value in supporting the diagnosis of H. pylori infection[24]. Therefore, IgA can be used for the diagnosis of H. pylori related dyspepsia. However, in the dyspeptic elderly patients with symptoms suggestive of ulcer and in those, who failed to respond to H2-receptor antagonists, endoscopy should be performed. The clinical significance of IgM antibodies, not tested in our study is debatable. Talley et al reported that levels of IgM and IgA antibodies are not consistently increased in peptic ulcer disease[27]. Lehn et al reported that detection of specific IgM does not seem to be of major value for adults and children[28]. In contrast, Andersen et al., have recommended inclusion of IgA and IgM antibodies to improve the diagnostic sensitivity of serology[20]. Another promising seroanalysis is detection of IgG antibody to H. pylori in urine using ELISA with a 95.9% sensitivity and 90% specificity[29]. Beside screening, serology was also found to be useful to monitor the success of therapy[30]. H. pylori IgG antibodies were found to decline after H. pylori eradication although this was not the purpose of our study.

In summary, our study supports the use of IgA and IgG for the diagnosis of H. pylori related dyspepsia. However, endoscopy should remain the method of choice especially in dyspeptic elderly patients not responding to treatment and for the diagnosis of duodenal and gastric ulcers.


   Acknowledgement   Top


We would like to express our thanks to the nursing staff in gastroenterology-endoscopy unit and technicians in Pathology Laboratories for their assistance and contribution to this study.

 
   References   Top

1. Warren JR. Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet 1983;1273.  Back to cited text no. 1    
2. Marshall B. Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet 1983;1273-4.  Back to cited text no. 2    
3. Parsonnet J, Hansen S, Rodriguez L, et al. Helicobacter pylori infection and gastric lymphoma. NEJM 1994;330:1267-70.  Back to cited text no. 3  [PUBMED]  [FULLTEXT]
4. Isaacson PG. Gastric Lymphoma and Helicobacter pylori. NEJM 1994;5:1310-11.  Back to cited text no. 4    
5. Satti MB, Twum-Danso K, Al-Freihi HM, et al. Helicobacter pylori-associated upper gastrointestinal disease in Saudi Arabia:a pathogenic evaluation of 298 endoscopic biopsies from 201 consecutive patients. J Gastroenterol 1990;85:527-34.  Back to cited text no. 5    
6. Mahmood K, Chowdhury MNH, Ayoola EA, Mofleh IA, Rashed RS, Faleh FZ. Helicobacterpylori:histiological and cultural correlation in chronic gastritis. Tropical Gastroenterol 1991;12:188-91.  Back to cited text no. 6    
7. Rashed RS, Ayoola EA, Mofleh IA, Chowdhury MN, Mahmood K, Faleh FZ. Helicobacter pylori and dyspepsia in an Arab population. Trop Georgr Med 1992;44:304-7.  Back to cited text no. 7    
8. Koestermann FK, Arsene D, Dalloul A. Helicobacter pylori antibodies in a Saudi population. Ann Saudi Med 1996;16:60-3.  Back to cited text no. 8    
9. Mohamed AE, Al-Karawi MA, Higham RH, et al. Comparison of Helicobacter serum. Saudi Med J 1997;18:74-7.  Back to cited text no. 9    
10. Booth L, Holdstock G, MacBride H, Hawtin P, et al. Clinical importance of Campylobacter pyloridis and associated serum IgG and IgA endoscopy. J Clin Pathol 1986;39:215-9.  Back to cited text no. 10  [PUBMED]  [FULLTEXT]
11. Jones DM, Eldridge J, Fox AJ, Sethi P, Whorwell PJ. Antibody to the gastric Campylobacter-like organism (Campylobacter pyloridis) clinical correlations and distribution in the normal population. J Med Microbiol 1986;22:57-62.  Back to cited text no. 11  [PUBMED]  
12. Graham DY, Kelin PD, Opekun AR, Bouton TW. Effect of age on the frequency of active Campylobacter pylori infection diagnosed by the [ 13 C] urea breath test in normal subjects and patients with peptic ulcer done. J Infect Dis 1988;157:777-80.  Back to cited text no. 12    
13. Buck GE. Campylobacter pylori and gastroduodenal disease. Clin Microbiol Rev 1990;3:1-12.  Back to cited text no. 13  [PUBMED]  [FULLTEXT]
14. Wilcox MH, Dent THS, Hunter JO, et al. Accuracy of serology for the diagnosis of Helicobacter pylori infection:a comparison of eight kits. J Clin Pathol 1996;49:373-6.  Back to cited text no. 14    
15. Abdullah AMA, Gad El Rab MO, Al Ayed 1, Kambal AM, Al Sohaibani, Al-Mazyad AS. Helicobacter pylori infection in children in Saudi Arabia. Trop Gastroenterol 1997;18:63-5.  Back to cited text no. 15    
16. Dhar R, Mustafa AS, Dhar PM, et al. Evaluation and comparison of two Immunodiagnostic assays for Helicobacter pylori antibodies with culture results. Diag Microbiol Infect Dis 1998;301:1-6.  Back to cited text no. 16    
17. Fendrick AM, Chemew ME, Hirth RA, Bloom BS. Alternative management strategies for patients with suspected peptic ulcer disease. Ann Intem Med 1995;123:260-8.  Back to cited text no. 17    
18. Thijs JC, Van Zwet AA, Thijs WJ, et al. Diagnostic tests for Helicobater pylori:a prospective evaluation of their accuracy, without selecting a single test as the gold standard. AM J Gastroenterol 1996;91:2125-9.  Back to cited text no. 18  [PUBMED]  
19. Sobala GM, Crabtree JE, Pentith JA, et al. Screening dyspepsia by serology to Helicobacterpylori. Lancet 1991;338:94-6.  Back to cited text no. 19  [PUBMED]  
20. Andersen LP, Rosenstock SJ, Bonnevie O, Jorgensen T. Seroprevalence of Immunoglobulin G,M and A antibodies to Helicobacter pylori in an unselected Danish population. Am J Epidem 1996;143:1157-64.  Back to cited text no. 20    
21. Briggs AH, Sculpher MJ, Logan RPH, Aldous J, Ramsay ME, Baron JH. Cost effectiveness of screening for and eradication of Helicobacter pylori in management of dyspeptic patient's under 45 years of age. BMJ 1996;312:1321-5.  Back to cited text no. 21    
22. Karvar S, Karch H, Frosch M, Burghardt W, Gross UWE. Use of serum-specific Immunoglobulin A and G for detection of Helicobacter pylori infection in patients with chronic gastritis by Immunosorbent analysis. J Clin Microbiol 1997;35:3058-61.  Back to cited text no. 22    
23. Azuma T, Kato T, Hirari M, Ito S, Kohli Y. Diagnosis of Helicobacter pylori infection. J Gastroenterol Hepatol 1996;11:662-9.  Back to cited text no. 23    
24. Jaskowski TD, Martins TB, Hill HR, Litwin CM. Immunoglobulin A antibodies to Helicobacter pylori. J Clin Microbiol 1997;35:2999-3000.  Back to cited text no. 24  [PUBMED]  [FULLTEXT]
25. Granberg C, Mansikka A, Lehtonen 0, et al. Diagnosis of Helicobacter pylori infection by using Pyloriset EIA-G and EIA-A for detection of serum Immunoglobulin G (IgG) and IgA antibodies. J Clin Microbiol 1993;31:1450-3.  Back to cited text no. 25    
26. Yamamoto L, Fukuda Y, Mizuta T, Fukada M, Nishigami T, Shimayama T. Serum anti-Helicobacter pylori antibodies and gastritis. J Clin Gastroenterol 1995;21:S164-8.  Back to cited text no. 26    
27. Talley NJ, Newell DG, Ormand JE. Serodiagnosis of Helicobacter pylori: comparison of enzyme-linked immunoassays. J Clin Microbiol 1991;29:1635-9.  Back to cited text no. 27    
28. Lehn N, Megraud F. Diagnosis of Helicobacter pylori. Curr Opinion Gastroenterol 12(Supp) 1996;6-10.  Back to cited text no. 28    
29. Alemohammed MM, Foley TJ, Cohen H. Detection of Immunoglobulin G antibodies to Helicobacter pylori in urine by an Enzyme Immunoassay Method. J Infect Dis 1993;31:2174-7.  Back to cited text no. 29    
30. Megraud F. Advantages and disadvantages of current diagnostic tests for the detection of Helicobacter pylori. Scand J Gastroenterol 1996;215 (Supp):57-62.  Back to cited text no. 30    

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Correspondence Address:
Hannan A.H Babay
Department of Pathology/Microbiology, (32), KKUH, P.O. Box 2925, Riyadh 11461
Saudi Arabia
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PMID: 19864726

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