Title: Ziehl Neelsen and Auramine Rhodamine staining: A comparative evaluation from North India

Authors: Kailash Chand Sharma, Ajay Yadav

 DOI: https://dx.doi.org/10.18535/jmscr/v8i6.117

Abstract

Introduction: Tubercular lymphadenitis can be presumptively diagnosed morphologically on fine-needle aspiration cytology of lymph node. Fine-needle aspiration cytology is now widely utilized as a first line diagnostic procedure in the diagnosis of palpable masses, including peripheral lymphadenopathy. Ziehl-Neelsen (ZN) method for acid-fast bacilli plays a key role in the diagnosis and also for the monitoring of treatment in tuberculosis. Its major disadvantage is low sensitivity ranging from 20% to 43%. Present study is to compare the efficacy of ZN staining method and fluorescent method using Auramine-Rhodamine in diagnosis of Tubercular Lymph Adenitis.

Methods: 100 samples were taken in this hospital based observational study. Patients with palpable swelling suspected to be tubercular in nature and confirmed by cytomorphology were included. Staining was done using Ziehl Neelsen and Auramine Rhodamine staining method. Comparison was made using grade for bacilli seen under the microscope.

Results: Maximum number of patients sample was seen in age group of 20-30 years. Female preponderance was seen. There was 59% samples where the aspirate was taken from the cervical lymph nodes. Maximum sample were seen of caseation with epitheloid cells followed by non-caseation with epitheloid cells. Positivity of aspirate was higher in staining by Auramine Rhodamine stain as compared to Ziehl Neelsen stain which was statistically significant.

Conclusion: The sensitivity of Auramine Rhodamine staining is higher which can be used in paucibacillary cases and it may be used in routine cases as well.

Keywords: Staining, Tuberculosis, Caseation, cytology.

References

  1. Dandapat MC, Mishra BM, Dash SP, Kar PK. Peripheral lymph node tuberculosis: A review of 80 cases. Br J Surg. 1990; 77(8):911-2.
  2. Lau SK, Kwan S, Lee J, Wei WI. Source of tubercle bacilli in cervical lymph nodes: a prospective study. J Laryngol Otol. 1991; 105(7):558-61.
  3. Daniel TM. Rapid diagnosis of tuberculosis: laboratory techniques applicable in developing countries. Rev Infect Dis. 1989; 2(Suppl 2):471-8.
  4. PG,Campbell PE. Tuberculous lymphadenitis in childhood: The significance of anonymous mycobacteria. Br J Surg 1962;50:302-14.
  5. Balows A, Hausler WJ, Hermann KL, Shadomy HJ. Manual of clinical Microbiology. 5th ed. Washington: D.C: American society for Microbiology 1991:308-11.
  6. Sharma SK, Mohan A. Extrapulmonary tuberculosis. Indian J Med Res 2004;120:316-53.
  7. Forbes BA, Sahm DF, Weissfeld AS. Mycobacteria. In: Bailey and Scotts′ Diagnostic Microbiology. 10 th Missouri: Mosby; 1988; p. 715-50.
  8. Thakur B, Mehrotra R, Nigam JS. Correlation of various techniques in diagnosis of tuberculous lymphadenitis on fine needle aspiration cytology. Patholog Res Int. 2013;2013:824620.
  9. Kumar N, Tiwari MC, Verma K. AFB staining in cytodiagnosis of tuberculosis without classical features: a comparison of Ziehl-Neelsen and fluorescent methods. Cytopathology. 1998;9(3):208-14.
  10. Wondwossen Rergette, Alemayehu Bekele. Acid-fast bacilli in aspiration smears from tuberculous patients. Ethiop J Health Dev. 2000,14(1) ; 99-104
  11. Annam V, Kulkarni MH, Puranik RB. Comparison of the modified fluorescent method and conventional Ziegl-Neelsen method in the detection of acid fast bacilli in lymph node aspirates. CytoJournal. 2009; 6:13-19.
  12. Fine needle aspiration cytology. Wikipedia, the free encyclopedia.http://en.wikipedia.org/wiki/Fine-needle_aspiration.
  13. Lau SK, Kwan S, Lee J, Wei WI. Source of tubercle bacilli in cervical lymph nodes: A prospective study. J Laryngol Otol 1991; 105:558-61
  14. Patra AK, Nanda BK, Mahapatra BK, Panda AK: Diagnosis of lymphadenopathy by fine needle aspiration biopsy. Indian J Pathol Microbiol. 1983; 26:273-78
  15. Rajwanshi A, Bhambhani S, Das DK. Fine needle aspiration cytology diagnosis of tuberculosis. Diagnostic Cytopathology 1987; 3: 13-16.
  16. Kumar N, Tiwari MC, Verma K. AFB staining in cytodiagnosis of tuberculosis without classical features: a comparison of Zeihl-Neelsen and fluorescent methods. Cytopathology 1998;9(3):208-14
  17. Metre MS, Jayaram G. Acid fast bacilli in aspiration smears from tuberculous lymph node. ActaCytol 1986; 31(1): 17-19
  18. Bailey TM, Akhtar M, Ali MA. Fine needle aspiration biopsy in the diagnosis of tuberculosis. Acta Cytol. 1985; 9:732-6
  19. Singh JP, Chaturvedi NK, Das A: Role of fine needle aspiration cytology in the diagnosis of tuberculous lymphadenitis. Indian J Pathol Microbiol. 1989; 32:101- 104.
  20. Nagpal BL, Dhar CN, Singh A, Bahl RA: Evaluation of imprint cytodiagnosis in cases of lymphadenopathy. Indian J Pathol Microbiol. 1982; 25:35-39.
  21. Truant JP, Brett WA and Thomas W, Jr. Fluorescence microscopy of tubercle bacilli stained with auramine and rhodamine. Henry Ford Hosp. Med. Bull. 1962; Vol. 10.
  22. Needham GM. Comparative efficacy of direct microscopy (two methods) and cultures in the diagnosis of tuberculosis. Proc. Staff Meet., Mayo Clinic. 1957; 32:1.
  23. Kuper SWA and May JR. Detection of acid fat organisms in tissue sections by fluorescence microscopy. J Path & Bact. 1960; 79: 59.
  24. Braunstein H and Adriano SM: Fluorescent stain for tubercle bacilli in histologic sections. Diagnostic efficiency in granulomatous lesions of lymph nodes. Am J Clin. Path. 1961; 36:37.
  25. Krishna M, Kumar A. Tuberculous mycobacteria bacilli fluorescence and compare with Ziehl- Neelsen stain in fine-needle aspiration cytology of tubercular lymph node. Int J Otorhinolaryngol Head Neck Surg. 2016;2:66-9.
  26. Dagar V, Heda S, Barsagade AK, Mahore S, Ambhore N. Comparision of ZN Staining and Fluorescent Microscopy in Detection of Acid Fast Bacilli in Fine Needle Aspiration Smears. IOSR Journal of Dental and Medical Sciences. 2016;15: 79-84.
  27. Laifangbam S, Singh HI, Singh NB, Devi KM, Singh NT. A Comparative Study of fluorescent microscopy with Ziehl-Neelsen staining and culture for the diagnosis of pulmonary tuberculosis. Kathmandu University Medical Journal. 2009; 7(27): 226-230
  28. Tulloch M. Tuberculosis. Blue Ridge Sanatorium http://www.faculty.virginia.edu/blueridgesanatorium

Corresponding Author

Kailash Chand Sharma

Department of Pathology, SMS Govt. Medical College & Hospital, Jaipur, India