Title: A Comparative study on staining for Helicobacter Pylori using new triple stain with routine Hematoxylin-Eosin and Giemsa stain

Authors: Krishna Sruthi R P, Preethamol S, Indira K

 DOI: https://dx.doi.org/10.18535/jmscr/v8i2.110

Abstract

   

Background: Helicobacter pylori, a spiral-shaped bacterium, can be seen in hematoxylin and eosin staining. However, when a low density of Helicobacter pylori and atrophic mucosal change are combined, it becomes difficult to demonstrate the organism. Hence many Pathologists now use Hematoxylin-Eosin plus a second stain for the bacteria. The aim of this study is to develop a New Triple Stain (Carbol Fuchsin/ Alcian Blue/ Hematoxylin-Eosin) is suitable for simultaneous visualization of Helicobacter pylori and gastric morphology.  

Objective: To introduce a New triple stain for simultaneous visualization of gastric morphology and Helicobacter pylori and compare it with routine Hematoxylin-Eosin and Giemsa.

Method: The study was carried out in Histopathology Laboratory and Dept. of MLT in Govt. Medical college Thiruvananthapuram. A total of 47 gastric biopsies confirmed as positive and negative by PCR were included in this study. Each sections from these biopsies stained with Hematoxylin-Eosin, Giemsa and New Triple stain and evaluated by the pathologist.

Result & Conclusion: The study was aimed to compare Hematoxylin-Eosin, Giemsa and New Triple for the detection of H. pylori. Morphological details are well observed in Hematoxylin-Eosin stain when compared to New Triple stain but bacterial demonstration is poor. Giemsa stain shows poor morphology when compared to both Hematoxylin-Eosin and New Triple Stain but bacterial identification is superior to Hematoxylin-Eosin. Hence it can only be used as a special stain for H.pylori. In histopathological aspect New Triple Stain shows greater sensitivity followed by Giemsa & Hematoxylin-Eosin.

Keywords:  PCR, Hematoxylin-Eosin, Giemsa, New Triple Stain.

References

  1. Eurogast Study Group. 1993. An international association between Helicobacter pylori infection and gastric cancer. Lancet, 341, 1359–62.
  2. Kasper D, Fauci A, Hauser S, Longo D, Jameson J, Loscalzo J. Harrison’s Principles of Internal Medicine. Star. 2015;2958.
  3. Vaira, D. and Vakil, N. 2001. Blood, urine, stool, breath, money and Helicobacter pylori. Gut, 48, 287–9.
  4. Madan, E., Kemp, J., et al. 1988. Evaluation of staining methods for identifying Campylobacter pylori. Am J ClinPathol, 90, 450–3.
  5. Hirschl, A.M., Rathbone, B.J., et al. 1990. Comparison of ELISA antigen preparations alone or in combination for serodiagnosing Helicobacter pylori infections. J ClinPathol, 43, 511–13.
  6. Braden B. Diagnosis of Helicobacter pylori infection. BMJ 2012; 344: e828 [PMID: 22368293 DOI: 10.1136/bmj.e828]
  7. Mamivand M, Shahreza S, Sharifi AH. Evaluation of methods for H. pylori detection in PPI consumption using culture, rapid urease test and smear examination. Ann Transl Med 2015; 3: 11 [PMID: 25705643 DOI: 10.3978/j.issn.2305-5839.2014.11.16]
  8. Diagnosis of Helicobacter pylori infection: Current options and developments, World J Gastroenterol 2015 October 28; 21(40): 11221-11235 ISSN 1007-9327 (print) ISSN 2219-2840 (online)
  9. Malfertheiner P, Megraud F, O’Morain CA, Atherton J, Axon AT, Bazzoli F, Gensini GF, Gisbert JP, Graham DY, Rokkas T, El-Omar EM, Kuipers EJ. Management of Helicobacter pylori infection—the Maastricht IV/ Florence Consensus Report. Gut 2012; 61: 646-664[PMID: 22491499 DOI: 10.1136/gutjnl-2012-302084].
  10. Madan E, Kemp J, Westblom TV, Subik M, Sexton S, Cook J. Evaluation of stain methods for identifying Campylobacter pylori. American Journal Clinical Pathology. 1988;90:450–454.

Corresponding Author

Preethamol S

Assistant Professor, Department of Pathology, Govt. Medical College, Thiruvananthapuram, Kerala