jsmc-10136

COMPARISON AMONG DIFFERENT LABORATORY TESTS FOR THE DIAGNOSIS OF HELICOBACTER PYLORI INFECTION IN DYSPEPTIC PATIENTS IN SULAIMANI GOVERNORATE  

Aryan Fuad H. Khorshid a, Sherko A Omer b and Aras Aziz Abdulla c 

a  Sulaimani Directorate of Health 

b Department of Microbiology, College of Medicine, University of Sulaimani.

c Department of Medicine, College of Medicine, University of Sulaimani.

Submitted: 11/9/2016; Accepted: 26/3/2017; Published: 21/12/2017

DOI Link: https://doi.org/10.17656/jsmc.10136

ABSTRACT

Background

Helicobacter pylori is a helical Gram-negative bacterium that colonizes human gastric mucosa leading to various pathologies including gastritis, gastric ulcers and gastric adenocarcinoma. Colonization is worldwide and affecting all age groups.

Objectives 

To identify H. pylori colonization among patients with dyspepsia using different laboratory tests including serology, stool antigen and PCR, and comparing the sensitivity and specificity of these tests.

Materials and Methods

From June through December 2014, one hundred patients with dyspepsia were enrolled in this study. They were 64 females (mean age 40.2 y) and 36 males (mean age 42.1 y). Two gastric antral biopsy specimens were taken in all patients submitted to esophagogastroduodenoscopy. Blood and stool specimens were also obtained from each patient. Four diagnostic laboratory tests were used for detection of the infection. Blood samples were tested for antibodies using rapid immunochromatography assay, stool for H. pylori antigen with immunochromatographic method and lastly PCR amplification of glmM gene was used to detect the organism in both gastric tissue and stool samples. 

Results

Esophagogastroduodenoscopy revealed that the majority of patients had gastritis and or duodenitis. Stool PCR for H. pylori was positive in 42% of the patients. Anti-H. pylori IgG in the blood and bacterial antigen were detected in 72% and 74% of patients respectively, while PCR test on gastric biopsy specimens was positive in 82% of the samples. 

Conclusion

H. pylori infection was detected regardless of the presence or absence of visible gastroduodenal pathology. PCR examination of gastric tissue was better than stool samples. In comparison to gastric biopsy PCR, stool antigen detection was found to be the most sensitive test reaching 87.8% while stool PCR was the most specific test. No single test solely can be used to diagnose or exclude colonization.

KEYWORDS

Diagnosis of H. pylori, Dyspepsia, Stool antigen, Antral biopsy, PCR.

References 

1. Andersen LP, Wadström T. Basic Bacteriology and Culture. In: Mobley HLT, Mendz GL, Hazell SL, editors. Helicobacter pylori: Physiology and Genetics. Washington (DC): ASM Press; 2001. Chapter 4. Available from: https://www.ncbi.nlm.nih.gov/books/NBK2444/.

2. Suerbaum S. The complex flagella of gastric Helicobacter species. Trends in microbiology. 1995;3(5):168-70; discussion 70-1.

3. Allaker RP, Young KA, Hardie JM, Domizio P, Meadows NJ. Prevalence of helicobacter pylori at oral and gastrointestinal sites in children: evidence for possible oral-to-oral transmission. Journal of medical microbiology. 2002;51(4):312-7.

4. Peach HG, Pearce DC, Farish SJ. Helicobacter pylori infection in an Australian regional city: prevalence and risk factors. The Medical journal of Australia. 1997;167(6):310-3.

5. Makristathis A, Pasching E, Schutze K, Wimmer M, Rotter ML, Hirschl AM. Detection of Helicobacter pylori in stool specimens by PCR and antigen enzyme immunoassay. Journal of clinical microbiology. 1998;36(9):2772-4.

6. Covacci A, Telford JL, Del Giudice G, Parsonnet J, Rappuoli R. Helicobacter pylori virulence and genetic geography. Science. 1999;284(5418):1328-33.

7. Schmitz A, Josenhans C, Suerbaum S. Cloning and characterization of the Helicobacter pylori flbA gene, which codes for a membrane protein involved in coordinated expression of flagellar genes. Journal of bacteriology. 1997;179(4):987-97.

8. Simon PM, Goode PL, Mobasseri A, Zopf D. Inhibition of Helicobacter pylori binding to gastrointestinal epithelial cells by sialic acid-containing oligosaccharides. Infection and immunity. 1997;65(2):750-7.

9. Ilver D, Arnqvist A, Ogren J, Frick IM, Kersulyte D, Incecik ET, et al. Helicobacter pylori adhesin binding fucosylated histo-blood group antigens revealed by retagging. Science. 1998;279(5349):373-7.

10. Bode G, Mauch F, Malfertheiner P. The coccoid forms of Helicobacter pylori. Criteria for their viability. Epidemiology and infection. 1993;111(3):483-90.

11. Harris PR, Mobley HL, Perez-Perez GI, Blaser MJ, Smith PD. Helicobacter pylori urease is a potent stimulus of mononuclear phagocyte activation and inflammatory cytokine production. Gastroenterology. 1996;111(2):419-25.

12. Cover TL, Blaser MJ. Purification and characterization of the vacuolating toxin from Helicobacter pylori. The Journal of biological chemistry. 1992;267(15):10570-5.

13. Ghiara P, Marchetti M, Blaser MJ, Tummuru MK, Cover TL, Segal ED, et al. Role of the Helicobacter pylori virulence factors vacuolating cytotoxin, CagA, and urease in a mouse model of disease. Infection and immunity. 1995;63(10):4154-60.

14. Walker MM, Crabtree JE. Helicobacter pylori infection and the pathogenesis of duodenal ulceration. Annals of the New York Academy of Sciences. 1998;859:96-111.

15. Crabtree JE, Kersulyte D, Li SD, Lindley IJ, Berg DE. Modulation of Helicobacter pylori induced interleukin-8 synthesis in gastric epithelial cells mediated by cag PAI encoded VirD4 homologue. Journal of clinical pathology. 1999;52(9):653-7.

16. Salzman A, Denenberg AG, Ueta I, O’Connor M, Linn SC, Szabo C. Induction and activity of nitric oxide synthase in cultured human intestinal epithelial monolayers. The American journal of physiology. 1996;270(4 Pt 1):G565-73.

17. Andrutis KA, Fox JG, Schauer DB, Marini RP, Murphy JC, Yan L, et al. Inability of an isogenic urease-negative mutant stain of Helicobacter mustelae to colonize the ferret stomach. Infection and immunity. 1995;63(9):3722-5.

18. Eaton KA, Krakowka S. Effect of gastric pH on urease-dependent colonization of gnotobiotic piglets by Helicobacter pylori. Infection and immunity. 1994;62(9):3604-7.

19. Harris PR, Smythies LE, Smith PD, Dubois A. Inflammatory cytokine mRNA expression during early and persistent Helicobacter pylori infection in nonhuman primates. The Journal of infectious diseases. 2000;181(2):783-6.

20. Blaser MJ, Berg DE. Helicobacter pylori genetic diversity and risk of human disease. The Journal of clinical investigation. 2001;107(7):767-73.

21. Dunn BE, Cohen H, Blaser MJ. Helicobacter pylori. Clinical microbiology reviews. 1997;10(4):720-41.

22. Kabir S. Detection of Helicobacter pylori in faeces by culture, PCR and enzyme immunoassay. Journal of medical microbiology. 2001;50(12):1021-9.

23. Malaty HM, Graham DY. Importance of childhood socioeconomic status on the current prevalence of Helicobacter pylori infection. Gut. 1994;35(6):742-5.

24. Ibrahim HAE, Al-Dhahir HAR, Mohammed MO. Effect of age, gender, blood group and social state on the sero-prevalence of Helicobacter pylori Infection among asymptomatic subjects in Sulaimani. Journal of Sulaimani Medical College. 2014;5(1).

25. Alsaimary I, Al-Sadoon M, Jassim A, Hamadi S. Clinical findings and Prevalence of Helicobacter pylori in Patients with Gastritis B in Al-basrah governorate. Oman medical journal. 2009;24(3):208-11.

26. Kalaf EA, Yassen NY, AL-Khafaji ZM, A’Rif GL, Abass BA. Multiplex PCR assay for detection of Helicobacter pylori isolated from Iraqi dyspeptic patients. Iraqi Journal of Cancer and Medical Genetics. 2013;6(1):49-54.

27. Megraud F, Lehours P. Helicobacter pylori detection and antimicrobial susceptibility testing. Clinical microbiology reviews. 2007;20(2):280-322.

28. Falsafi T, Favaedi R, Mahjoub F, Najafi M. Application of stool-PCR test for diagnosis of Helicobacter pylori infection in children. World journal of gastroenterology : WJG. 2009;15(4):484-8.

29. Megraud F, Broutet N. Review article: have we found the source of Helicobacter pylori? Alimentary pharmacology & therapeutics. 2000;14 Suppl 3:7-12.

30. Redeen S, Petersson F, Jonsson KA, Borch K. Relationship of gastroscopic features to histological findings in gastritis and Helicobacter pylori infection in a general population sample. Endoscopy. 2003;35(11):946-50.

31. Herbrink P, van Doorn LJ. Serological methods for diagnosis of Helicobacter pylori infection and monitoring of eradication therapy. European journal of clinical microbiology & infectious diseases : official publication of the European Society of Clinical Microbiology. 2000;19(3):164-73.

32. Anderson JC, Cheng E, Roeske M, Marchildon P, Peacock J, Shaw RD. Detection of serum antibodies to Helicobacter pylori by an immunochromatographic method. The American journal of gastroenterology. 1997;92(7):1135-9.

33. Gisbert JP, de la Morena F, Abraira V. Accuracy of monoclonal stool antigen test for the diagnosis of H. pylori infection: a systematic review and meta-analysis. The American journal of gastroenterology. 2006;101(8):1921-30.

34. Simala-Grant JL, Taylor DE. Molecular biology methods for the characterization of Helicobacter pylori infections and their diagnosis. APMIS : acta pathologica, microbiologica, et immunologica Scandinavica. 2004;112(11-12):886-97.

35. Kisa O, Albay A, Mas MR, Celasun B, Doganci L. The evaluation of diagnostic methods for the detection of Helicobacter pylori in gastric biopsy specimens. Diagnostic microbiology and infectious disease. 2002;43(4):251-5.

36. Chisholm SA, Owen RJ. Application of polymerase chain reaction-based assays for rapid identification and antibiotic resistance screening of Helicobacter pylori in gastric biopsies. Diagnostic microbiology and infectious disease. 2008;61(1):67-71.

37. Thoreson AC, Borre M, Andersen LP, Jorgensen F, Kiilerich S, Scheibel J, et al. Helicobacter pylori detection in human biopsies: a competitive PCR assay with internal control reveals false results. FEMS immunology and medical microbiology. 1999;24(2):201-8.

38. Kansau I, Raymond J, Bingen E, Courcoux P, Kalach N, Bergeret M, et al. Genotyping of Helicobacter pylori isolates by sequencing of PCR products and comparison with the RAPD technique. Research in microbiology. 1996;147(8):661-9.

39. Pajares-Garcia JM. Diagnosis of Helicobacter pylori: invasive methods. Italian journal of gastroenterology and hepatology. 1998;30 Suppl 3:S320-3.

40. Liao CC, Lee CL, Lai YC, Huang SH, Lee SC, Wu CH, et al. Accuracy of three diagnostic tests used alone and in combination for detecting Helicobacter pylori infection in patients with bleeding gastric ulcers. Chinese medical journal. 2003;116(12):1821-6.

41. Hino B, Eliakim R, Levine A, Sprecher H, Berkowitz D, Hartman C, et al. Comparison of invasive and non-invasive tests diagnosis and monitoring of Helicobacter pylori infection in children. Journal of pediatric gastroenterology and nutrition. 2004;39(5):519-23.

42. McNulty CA, Nair P, Watson BE, Uff JS, Valori RM. A comparison of six commercial kits for Helicobacter pylori detection. Communicable disease and public health / PHLS. 1999;2(1):59-63.

43. Feldman M, Cryer B, Lee E, Peterson WL. Role of seroconversion in confirming cure of Helicobacter pylori infection. Jama. 1998;280(4):363-5.

44. Andrews J, Marsden B, Brown D, Wong VS, Wood E, Kelsey M. Comparison of three stool antigen tests for Helicobacter pylori detection. Journal of clinical pathology. 2003;56(10):769-71.

45. Brooks HJ, Ahmed D, McConnell MA, Barbezat GO. Diagnosis of helicobacter pylori infection by polymerase chain reaction: is it worth it? Diagnostic microbiology and infectious disease. 2004;50(1):1-5.

46. Al-Dhaher Z. Study of Some Bacteriological and Immunological Aspects of H. pylori Al-Mustansiriya University; 2001.

47. Mohammed O. Mohammed. Correlation of Endoscopic Findings with Various Helicobacter pylori Tests among Dyspeptic Patients. International Journal of Clinical Medicine. 2014;5:1180-8 

48. Macarthur C, Saunders N, Feldman W. Helicobacter pylori, gastroduodenal disease, and recurrent abdominal pain in children. Jama. 1995;273(9):729-34.

49. Klein PD, Gilman RH, Leon-Barua R, Diaz F, Smith EO, Graham DY. The epidemiology of Helicobacter pylori in Peruvian children between 6 and 30 months of age. The American journal of gastroenterology. 1994;89(12):2196-200.

50. Ma JL, You WC, Gail MH, Zhang L, Blot WJ, Chang YS, et al. Helicobacter pylori infection and mode of transmission in a population at high risk of stomach cancer. International journal of epidemiology. 1998;27(4):570-3.

51. Rauws EA, Tytgat GN. Cure of duodenal ulcer associated with eradication of Helicobacter pylori. Lancet. 1990;335(8700):1233-5.

52. Wroblewski LE, Peek RM, Jr., Wilson KT. Helicobacter pylori and gastric cancer: factors that modulate disease risk. Clinical microbiology reviews. 2010;23(4):713-39.

53. Cognein P, Costa A, Giacosa A. Serodiagnosis of Helicobacter pylori: evaluation of a rapid, miniaturized immunochromatographic test. European journal of cancer prevention : the official journal of the European Cancer Prevention Organisation. 1994;3(6):457-63.

54. Eugenia M, Quintana-Guzman, Schosinky-Nevermann K, Arias-Echandi ML, Davidovich-Rose H. Comparative study of urease tests for Helicobacter pylori detection In gastric biopsies. Rev Biomed. 1999;10:145-51.

55. Baqai R, Qureshi H, Arian G, Mehdi I. Diagnostic efficacy of stool antigen test (HPSA), CLO test and serology for the detection of Helicobacter pylori infection. Journal of Ayub Medical College, Abbottabad : JAMC. 2003;15(4):34-6.

56. Li YH, Guo H, Zhang PB, Zhao XY, Da SP. Clinical value of Helicobacter pylori stool antigen test, ImmunoCard STAT HpSA, for detecting H pylori infection. World journal of gastroenterology : WJG. 2004;10(6):913-4.

57. Al-Yas M. Comparative Study Between H. pylori Isolated from Human and Some Domestic Animals: Al-Nahrain University; 2006.

58. Twaij A. Invasive and Non Invasive Methods for Detection of H. pylori With Some Molecular Aspects of its Pathogenesis [PhD Thesis]: Al-Nahrain University; 2006.

59. Lu JJ, Perng CL, Shyu RY, Chen CH, Lou Q, Chong SK, et al. Comparison of five PCR methods for detection of Helicobacter pylori DNA in gastric tissues. Journal of clinical microbiology. 1999;37(3):772-4.

60. Patel SK, Pratap CB, Jain AK, Gulati AK, Nath G. Diagnosis of Helicobacter pylori: what should be the gold standard? World journal of gastroenterology : WJG. 2014;20(36):12847-59.

61. Mishra S, Singh V, Rao GR, Jain AK, Dixit VK, Gulati AK, et al. Detection of Helicobacter pylori in stool specimens: comparative evaluation of nested PCR and antigen detection. Journal of infection in developing countries. 2008;2(3):206-10.

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