Generic selectors
Exact matches only
Search in title
Search in content
Post Type Selectors
Search in posts
Search in pages
Filter by Categories
15th National Conference of the IAOMFP, Chennai, 2006
Abstract
Abstracts from current literature
Acne in India: Guidelines for management - IAA Consensus Document
Addendum
Announcement
Art & Psychiatry
Article
Articles
Association Activities
Association Notes
Award Article
Book Review
Brief Report
Case Analysis
Case Letter
Case Letters
Case Notes
Case Report
Case Reports
Clinical and Laboratory Investigations
Clinical Article
Clinical Studies
Clinical Study
Commentary
Conference Oration
Conference Summary
Continuing Medical Education
Correspondence
Corrigendum
Cosmetic Dermatology
Cosmetology
Current Best Evidence
Current Issue
Current View
Derma Quest
Dermato Surgery
Dermatopathology
Dermatosurgery Specials
Dispensing Pearl
Do you know?
Drug Dialogues
e-IJDVL
Editor Speaks
Editorial
Editorial Remarks
Editorial Report
Editorial Report - 2007
Editorial report for 2004-2005
Errata
Erratum
Focus
Fourth All India Conference Programme
From Our Book Shelf
From the Desk of Chief Editor
General
Get Set for Net
Get set for the net
Guest Article
Guest Editorial
History
How I Manage?
IADVL Announcement
IADVL Announcements
IJDVL Awards
IJDVL AWARDS 2015
IJDVL Awards 2018
IJDVL Awards 2019
IJDVL Awards 2020
IJDVL International Awards 2018
Images in Clinical Practice
Images in Dermatology
In Memorium
Inaugural Address
Index
Knowledge From World Contemporaries
Leprosy Section
Letter in Response to Previous Publication
Letter to Editor
Letter to the Editor
Letter to the Editor - Case Letter
Letter to the Editor - Letter in Response to Published Article
LETTER TO THE EDITOR - LETTERS IN RESPONSE TO PUBLISHED ARTICLES
Letter to the Editor - Observation Letter
Letter to the Editor - Study Letter
Letter to the Editor - Therapy Letter
Letter to the Editor: Articles in Response to Previously Published Articles
Letters in Response to Previous Publication
Letters to the Editor
Letters to the Editor - Letter in Response to Previously Published Articles
Letters to the Editor: Case Letters
Letters to the Editor: Letters in Response to Previously Published Articles
Media and news
Medicolegal Window
Messages
Miscellaneous Letter
Musings
Net Case
Net case report
Net Image
Net Images
Net Letter
Net Quiz
Net Study
New Preparations
News
News & Views
Obituary
Observation Letter
Observation Letters
Oration
Original Article
ORIGINAL CONTRIBUTION
Original Contributions
Pattern of Skin Diseases
Pearls
Pediatric Dermatology
Pediatric Rounds
Perspective
Presedential Address
Presidential Address
Presidents Remarks
Quiz
Recommendations
Regret
Report
Report of chief editor
Report of Hon : Treasurer IADVL
Report of Hon. General Secretary IADVL
Research Methdology
Research Methodology
Resident page
Resident's Page
Resident’s Page
Residents' Corner
Residents' Corner
Residents' Page
Retraction
Review
Review Article
Review Articles
Reviewers 2022
Revision Corner
Self Assessment Programme
SEMINAR
Seminar: Chronic Arsenicosis in India
Seminar: HIV Infection
Short Communication
Short Communications
Short Report
Snippets
Special Article
Specialty Interface
Studies
Study Letter
Study Letters
Supplement-Photoprotection
Supplement-Psoriasis
Symposium - Contact Dermatitis
Symposium - Lasers
Symposium - Pediatric Dermatoses
Symposium - Psoriasis
Symposium - Vesicobullous Disorders
SYMPOSIUM - VITILIGO
Symposium Aesthetic Surgery
Symposium Dermatopathology
Symposium-Hair Disorders
Symposium-Nails Part I
Symposium-Nails-Part II
Systematic Review and Meta-Analysis
Systematic Reviews and Meta-analyses
Systematic Reviews and Meta-analysis
Tables
Technology
Therapeutic Guideline-IADVL
Therapeutic Guidelines
Therapeutic Guidelines - IADVL
Therapeutics
Therapy
Therapy Letter
Therapy Letters
View Point
Viewpoint
What’s new in Dermatology
Generic selectors
Exact matches only
Search in title
Search in content
Post Type Selectors
Search in posts
Search in pages
Filter by Categories
15th National Conference of the IAOMFP, Chennai, 2006
Abstract
Abstracts from current literature
Acne in India: Guidelines for management - IAA Consensus Document
Addendum
Announcement
Art & Psychiatry
Article
Articles
Association Activities
Association Notes
Award Article
Book Review
Brief Report
Case Analysis
Case Letter
Case Letters
Case Notes
Case Report
Case Reports
Clinical and Laboratory Investigations
Clinical Article
Clinical Studies
Clinical Study
Commentary
Conference Oration
Conference Summary
Continuing Medical Education
Correspondence
Corrigendum
Cosmetic Dermatology
Cosmetology
Current Best Evidence
Current Issue
Current View
Derma Quest
Dermato Surgery
Dermatopathology
Dermatosurgery Specials
Dispensing Pearl
Do you know?
Drug Dialogues
e-IJDVL
Editor Speaks
Editorial
Editorial Remarks
Editorial Report
Editorial Report - 2007
Editorial report for 2004-2005
Errata
Erratum
Focus
Fourth All India Conference Programme
From Our Book Shelf
From the Desk of Chief Editor
General
Get Set for Net
Get set for the net
Guest Article
Guest Editorial
History
How I Manage?
IADVL Announcement
IADVL Announcements
IJDVL Awards
IJDVL AWARDS 2015
IJDVL Awards 2018
IJDVL Awards 2019
IJDVL Awards 2020
IJDVL International Awards 2018
Images in Clinical Practice
Images in Dermatology
In Memorium
Inaugural Address
Index
Knowledge From World Contemporaries
Leprosy Section
Letter in Response to Previous Publication
Letter to Editor
Letter to the Editor
Letter to the Editor - Case Letter
Letter to the Editor - Letter in Response to Published Article
LETTER TO THE EDITOR - LETTERS IN RESPONSE TO PUBLISHED ARTICLES
Letter to the Editor - Observation Letter
Letter to the Editor - Study Letter
Letter to the Editor - Therapy Letter
Letter to the Editor: Articles in Response to Previously Published Articles
Letters in Response to Previous Publication
Letters to the Editor
Letters to the Editor - Letter in Response to Previously Published Articles
Letters to the Editor: Case Letters
Letters to the Editor: Letters in Response to Previously Published Articles
Media and news
Medicolegal Window
Messages
Miscellaneous Letter
Musings
Net Case
Net case report
Net Image
Net Images
Net Letter
Net Quiz
Net Study
New Preparations
News
News & Views
Obituary
Observation Letter
Observation Letters
Oration
Original Article
ORIGINAL CONTRIBUTION
Original Contributions
Pattern of Skin Diseases
Pearls
Pediatric Dermatology
Pediatric Rounds
Perspective
Presedential Address
Presidential Address
Presidents Remarks
Quiz
Recommendations
Regret
Report
Report of chief editor
Report of Hon : Treasurer IADVL
Report of Hon. General Secretary IADVL
Research Methdology
Research Methodology
Resident page
Resident's Page
Resident’s Page
Residents' Corner
Residents' Corner
Residents' Page
Retraction
Review
Review Article
Review Articles
Reviewers 2022
Revision Corner
Self Assessment Programme
SEMINAR
Seminar: Chronic Arsenicosis in India
Seminar: HIV Infection
Short Communication
Short Communications
Short Report
Snippets
Special Article
Specialty Interface
Studies
Study Letter
Study Letters
Supplement-Photoprotection
Supplement-Psoriasis
Symposium - Contact Dermatitis
Symposium - Lasers
Symposium - Pediatric Dermatoses
Symposium - Psoriasis
Symposium - Vesicobullous Disorders
SYMPOSIUM - VITILIGO
Symposium Aesthetic Surgery
Symposium Dermatopathology
Symposium-Hair Disorders
Symposium-Nails Part I
Symposium-Nails-Part II
Systematic Review and Meta-Analysis
Systematic Reviews and Meta-analyses
Systematic Reviews and Meta-analysis
Tables
Technology
Therapeutic Guideline-IADVL
Therapeutic Guidelines
Therapeutic Guidelines - IADVL
Therapeutics
Therapy
Therapy Letter
Therapy Letters
View Point
Viewpoint
What’s new in Dermatology

Translate this page into:

Original Article
2003:69:2;173-174
PMID: 17642871

Role of modified rapid AFB method in histopathological sections of hansen's disease

SV Nayak, AS Shivarudrappa, AH Nagarajappa, S Sacchidanand, SM Ahmed
 Dept. of Pathology & Dermatology, Victoria Hospital, Banglore, India

Correspondence Address:
S V Nayak
H No. 4, "Prubhu Nilaya", 3rd Cross, Subbanna Gerden, Vijaynagar, Banglore - 560 040, Karnataka
India
How to cite this article:
Nayak S V, Shivarudrappa A S, Nagarajappa A H, Sacchidanand S, Ahmed S M. Role of modified rapid AFB method in histopathological sections of hansen's disease. Indian J Dermatol Venereol Leprol 2003;69:173-174
Copyright: (C)2003 Indian Journal of Dermatology, Venereology, and Leprology

Abstract

Leprosy is a chronic debilitating disease. A reliable diagnosis hinges around a good histopothological diagnosis and demonstration of the bacilli in the histopathological section. The usual method performed Modified Fite Faraco Method is time consuming, laborious and less sensitive. It has been our endeavor to devise a more rapid and sensitive method for the diagnosis and bacillary load detection in histopathological sections. The Modified Rapid AFB devised by us is sensitive and time saving which is essential for the pathologist and for the treatment by the Dermatologist. We hove studied about 53 cases of different types of Hansens disease and compared with both Modified Fite Faraco method and Modified Rapid AFB method. The results were found to be very encouraging with the Modified Rapid AFB method.
Keywords: Hansen′s disease, leprosy, Fite Faraco method, Modified Rapid AFB method

Introduction

Modifed Fite Faraco is the oldest method used to detect Mycobacterium leprae in tissues.[1],[2] Pararosanaline stain has been used in some studies to detect Mycobacterium leprae and was found to be sensitive in few studies.[3],[1] It was our endeavor to devise a new method to increase the sensitivity of detecting Mycobacterium leprae in tissue sections by using pararosanaline stain and periodic acid especially for paucibacillary cases. We describe our experience with Modified Fite Faraco and Modified Rapid AFB Method in histopathological sections.

Materials and Methods

Fifty-three leprosy cases attending Victoria Hospital and Bowring and Lady Curzon Hospital from Mar 2001 to Sept 2002, Bangalore were involved in the study. Punch biopsies were taken from the active lesion and classified according to clinical features, slit-skin smear study, histopathological features and Modified Fite Faraco method into: Indeterminate leprosy, tuberculoid leprosy, borderline tuberculoid leprosy, mid borderline leprosy, borderline lepromatous leprosy and lepromatous leprosy.

The biopsies were routinely processed. Haematoxylin eosin stained sections were studied. Ribbons containing 10 serial sections, 5 pm thick were taken, 5 each for Modified Fite Faraco method and 5 for Modified Rapid AFB method.

Modified Rapid AFB method

Ribbons containing 5 serial sections, 5 Nm thick were taken on a clean glass slide with egg albumin.

1. Deparaffinization was done by 1:3 peanut oil Xylene.

2. Blot dry

3. Dip slide in 10% periodic acid for 30 minutes.

4. Dip it in water for 2-3 times

5. Blot dry

6. Flood the slide with pararosanaline stain and keep it in the incubator at 70 Degrees for 8-10 minutes.

7. Wash in running tap water.

8. Blot dry.

9. Decolorise with 1 % hydrocloric acid in 70% ethanol until faint pink.

10. Wash it in running tap water.

11. Blot dry.

12. Counterstain with 0.1% malachite green or 1 methylene blue for 30 seconds to 1 minute

13. Wash it in running tap water.

14. Blot dry.

15. Dehydrate by dipping in absolute alcohol (1-2 dips).

16. Blot dry immediately.

17. Dip in xylene and mount in DPX immediately.

The slides were then screened with 1 Ox, 40x and 100x objective and findings noted.

Modified Fite Faraco method

Ribbons containing 5 serial sections were taken on a clean glass slide with egg albumin and stained conventionally, and the findings noted.

Discussion

The conventional Fite Faraco method is the most commonest method used for tissue sections in detecting Mycobacterium leprae and has stood the test of time. In our study the combination of periodic acid and pararosanaline stain was found to be very effective. The sensitivity with Modified Rapid AFB method was found to be 54.7%.

The com-bination of pe-riodic acid and pararosanaline stain can be used as a supplement or can be used per se in paucibacillary cases.

We faced few technical problems during this procedure such as sections getting detached from the slide while staining. This was overcome by hav-ing thin sections.

The sections should be absolutely thin and complete deparaffinisation of the section should be done otherwise it effects the process of decolorisation.

Clinical correlation was carried out on all cases. Out of 54 cases diagnosed histopathologicaly, in 39 cases clinical and histopathological correlation was established. The remaining 14 cases mostly belonging to paucibacillary type, no histopathological or clinical correlation could be established. It is in these cases that modified rapid AFB method plays a role in typing of leprosy.

The coincidence of classification by histopathological and clinicial examination as percentage for each mode is shown in [Table - 2]. Maximum clinical correlation was seen in lepromatous leprosy (100%) and midborderline (100%) were as clinical cor-relation was found to be lacking in border-line tuberculoid leprosy (76.92%), tubercu-loid leprosy (66.66%). The clinical correla-tion was least in indeterminate leprosy (63.64%). The overall coincidence of di-agnosis of classification was seen in 39 cases (73.55%) study.

We agree with Prakash Kumar[5] et al that maxium clinical correlation can be established in multibacillary type of leprosy than in paucibacillary type of leprosy.

References
1.
JOB CK, and Chocko CJG. Modification of Fite's stain for demonstration of M. leprae in tissue sections. Indian J Lepr,1986; 58: 17-19.
[Google Scholar]
2.
Samuel KC, Chatterjee SN. Modification of Fite Faroco staining for acid fast bacilli, Indian J Path Badl971; 14: 107-109.
[Google Scholar]
3.
Harada K, Suzuki K. Improved staining of leprosy bacilli in tissues. Lepr Rev 1989; 60:124-128.
[Google Scholar]
4.
Harada K. A modified allochorome procedure for demonstrating mycobacteria in tissue sections, Int J Lepr 1976; 5:49-51.
[Google Scholar]
5.
Nironjona Murthy B, Prakash Kumar, KR Chatura, Chondrashekar Basavaraj. Histopathological correlation of skin biopsies in leprosy. Indian J Dermatol Venereal Leprol 2001; 61: 299-301.
[Google Scholar]
Show Sections