Evaluation of fluorescent staining for diagnosis of leprosy and its impact on grading of the disease

Comparison with conventional staining

Deepa Sowkur Anandarama Adiga, Surekha B. Hippargi, Gururaja Rao, Debarshi Saha, Bheemrao Ramling Yelikar, Mahesh Karigoudar

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Introduction: Leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae). Histopathological examination of skin lesion is the gold standard for diagnosis. We evaluated the possible role of fluorescent microscopy in this direction which is increasingly used for rapid screening. Aim: To compare the efficacy of auramine rhodamine stain with Ziehl-Neelsen and modified Fite-faraco staining in diagnosing M. leprae in tissue sections. Study Design: Experimental, cross-sectional and retrospective study conducted for 4 years. Methods and Materials: Skin biopsies of sixty clinically diagnosed leprosy patients were stained by Ziehl-Neelsen, Fite-Faraco and fluorescent stain. The presence of the bacilli and the bacillary index was scored for each case. The bacillary index by each staining methods were compared. Statistical Analysis: SPSS v 17 (IBM, New York) used for data analysis. Chi-Square test was used to calculate significance between differences. The p-value of <0.05 was considered as statistically significant. Pearson Correlation (r-value determined) was also used for comparison between groups. Results: Sensitivity of fluorescent stain for indeterminate and borderline tuberculoid leprosies were 100% each. Positivity rates and mean bacteriological index with fluorescent stain was higher (43.3 and 11.5 respectively) as compared to that of Ziehl- Neelsen and Fite-faraco when the bacillary load was less (bacillary index < 3). There was significant correlation between the three staining types at higher bacillary load. There was a higher mean bacillary index with fluorescent stain as well as detection of an additional multibacillary case. Conclusion: Fluorescent method is more sensitive than modified fite-faraco method in detecting lepra bacilli in tissue sections especially in cases with bacillary index less than three. With its higher sensitivity, paucibacillary cases could be upgraded to multibacillary thus affecting treatement decisions.

Original languageEnglish
Pages (from-to)EC23-EC26
JournalJournal of Clinical and Diagnostic Research
Volume10
Issue number10
DOIs
Publication statusPublished - 01-10-2016

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Leprosy
Coloring Agents
Staining and Labeling
Paucibacillary Leprosy
Mycobacterium leprae
Bacilli
Bacillus
Skin
Benzophenoneidum
Tissue
Rhodamines
Biopsy
Chi-Square Distribution
Design of experiments
Communicable Diseases
Microscopy
Statistical methods
Microscopic examination
Screening
Chronic Disease

All Science Journal Classification (ASJC) codes

  • Clinical Biochemistry

Cite this

Adiga, Deepa Sowkur Anandarama ; Hippargi, Surekha B. ; Rao, Gururaja ; Saha, Debarshi ; Yelikar, Bheemrao Ramling ; Karigoudar, Mahesh. / Evaluation of fluorescent staining for diagnosis of leprosy and its impact on grading of the disease : Comparison with conventional staining. In: Journal of Clinical and Diagnostic Research. 2016 ; Vol. 10, No. 10. pp. EC23-EC26.
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Evaluation of fluorescent staining for diagnosis of leprosy and its impact on grading of the disease : Comparison with conventional staining. / Adiga, Deepa Sowkur Anandarama; Hippargi, Surekha B.; Rao, Gururaja; Saha, Debarshi; Yelikar, Bheemrao Ramling; Karigoudar, Mahesh.

In: Journal of Clinical and Diagnostic Research, Vol. 10, No. 10, 01.10.2016, p. EC23-EC26.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Evaluation of fluorescent staining for diagnosis of leprosy and its impact on grading of the disease

T2 - Comparison with conventional staining

AU - Adiga, Deepa Sowkur Anandarama

AU - Hippargi, Surekha B.

AU - Rao, Gururaja

AU - Saha, Debarshi

AU - Yelikar, Bheemrao Ramling

AU - Karigoudar, Mahesh

PY - 2016/10/1

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N2 - Introduction: Leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae). Histopathological examination of skin lesion is the gold standard for diagnosis. We evaluated the possible role of fluorescent microscopy in this direction which is increasingly used for rapid screening. Aim: To compare the efficacy of auramine rhodamine stain with Ziehl-Neelsen and modified Fite-faraco staining in diagnosing M. leprae in tissue sections. Study Design: Experimental, cross-sectional and retrospective study conducted for 4 years. Methods and Materials: Skin biopsies of sixty clinically diagnosed leprosy patients were stained by Ziehl-Neelsen, Fite-Faraco and fluorescent stain. The presence of the bacilli and the bacillary index was scored for each case. The bacillary index by each staining methods were compared. Statistical Analysis: SPSS v 17 (IBM, New York) used for data analysis. Chi-Square test was used to calculate significance between differences. The p-value of <0.05 was considered as statistically significant. Pearson Correlation (r-value determined) was also used for comparison between groups. Results: Sensitivity of fluorescent stain for indeterminate and borderline tuberculoid leprosies were 100% each. Positivity rates and mean bacteriological index with fluorescent stain was higher (43.3 and 11.5 respectively) as compared to that of Ziehl- Neelsen and Fite-faraco when the bacillary load was less (bacillary index < 3). There was significant correlation between the three staining types at higher bacillary load. There was a higher mean bacillary index with fluorescent stain as well as detection of an additional multibacillary case. Conclusion: Fluorescent method is more sensitive than modified fite-faraco method in detecting lepra bacilli in tissue sections especially in cases with bacillary index less than three. With its higher sensitivity, paucibacillary cases could be upgraded to multibacillary thus affecting treatement decisions.

AB - Introduction: Leprosy is a chronic infectious disease caused by Mycobacterium leprae (M. leprae). Histopathological examination of skin lesion is the gold standard for diagnosis. We evaluated the possible role of fluorescent microscopy in this direction which is increasingly used for rapid screening. Aim: To compare the efficacy of auramine rhodamine stain with Ziehl-Neelsen and modified Fite-faraco staining in diagnosing M. leprae in tissue sections. Study Design: Experimental, cross-sectional and retrospective study conducted for 4 years. Methods and Materials: Skin biopsies of sixty clinically diagnosed leprosy patients were stained by Ziehl-Neelsen, Fite-Faraco and fluorescent stain. The presence of the bacilli and the bacillary index was scored for each case. The bacillary index by each staining methods were compared. Statistical Analysis: SPSS v 17 (IBM, New York) used for data analysis. Chi-Square test was used to calculate significance between differences. The p-value of <0.05 was considered as statistically significant. Pearson Correlation (r-value determined) was also used for comparison between groups. Results: Sensitivity of fluorescent stain for indeterminate and borderline tuberculoid leprosies were 100% each. Positivity rates and mean bacteriological index with fluorescent stain was higher (43.3 and 11.5 respectively) as compared to that of Ziehl- Neelsen and Fite-faraco when the bacillary load was less (bacillary index < 3). There was significant correlation between the three staining types at higher bacillary load. There was a higher mean bacillary index with fluorescent stain as well as detection of an additional multibacillary case. Conclusion: Fluorescent method is more sensitive than modified fite-faraco method in detecting lepra bacilli in tissue sections especially in cases with bacillary index less than three. With its higher sensitivity, paucibacillary cases could be upgraded to multibacillary thus affecting treatement decisions.

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