Generic selectors
Exact matches only
Search in title
Search in content
Post Type Selectors
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
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
Medicolegal Window
Messages
Miscellaneous Letter
Musings
Net Case
Net case report
Net Image
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
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 Reviews and Meta-analyses
Systematic Reviews and Meta-analysis
Tables
Technology
Therapeutic Guideline-IADVL
Therapeutic Guidelines
Therapeutic Guidelines - IADVL
Therapeutics
Therapy
Therapy Letter
View Point
Viewpoint
What’s new in Dermatology
View/Download PDF

Translate this page into:

Observation Letter
ARTICLE IN PRESS
doi:
10.25259/IJDVL_581_2021

Masson-Fontana stain: A silver lining for diagnosis of primary syphilitic chancre

Department of Dermatology, Amrita Institute of Medical Sciences, Amrita Vishwa Vidyapeetham, Kochi, Kerala, India
Department of Microbiology, Amrita Institute of Medical Sciences, Amrita Vishwa Vidyapeetham, Kochi, Kerala, India
Corresponding Author: Dr. Gopikrishnan Anjaneyan, Department of Dermatology, Amrita Institute of Medical Sciences, Amrita Vishwa Vidyapeetham, Ponekkara, Kochi, Kerala, India. drgopikrishnana@gmail.com
Licence
This is an open-access article distributed under the terms of the Creative Commons Attribution-Non Commercial-Share Alike 4.0 License, which allows others to remix, transform, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.

How to cite this article: Anjaneyan G, Kumar A, Thomas J. Masson-Fontana stain: A silver lining for diagnosis of primary syphilitic chancre. Indian J Dermatol Venereol Leprol doi: 10.25259/IJDVL_581_2021

Sir,

Syphilis is a sexually transmitted infection caused by the spirochaete, Treponema pallidum subsp. Pallidum. Its first stage, primary syphilis, is a local infection due to spirochaete replication at the site of inoculation, after contact with an infected person.

Treponema pallidum is a motile, slow-growing fastidious bacterium of the order Spirochaetales and humans are their only natural host. It is a slender spiral bacterium measuring around 10–15 microns in length and 0.10–0.15 microns in width, and thus, difficult to visualize on direct microscopy.1 We successfully demonstrated spirochaetes by light microscopic examination from Masson-Fontana silver-stained smear from a suspected syphilitic chancre.

A 30-year-old unmarried male presented with a single painless penile ulcer of one-week duration. He reported sexual contact with three different partners, with the last unprotected exposure occurring two months back. Examination revealed a single indurated ulcer sized 2×1 cm at the coronal sulcus [Figure 1] with two discrete non-tender right inguinal lymph nodes. There were no systemic complaints and examination of other sites including oral and perianal areas were unremarkable. Serological tests for other sexually transmitted infections including HIV, hepatitis B and C were negative.

Figure 1:: Single hard chancre at the coronal sulcus

Rapid plasma reagin was reactive only in undiluted serum and not in serial dilutions. To exclude false positivity, we sent the serum for Treponema pallidum haemagglutination test along with a smear from ulcer base for Masson-Fontana silver staining. The Masson-Fontana silver stained smear demonstrated multiple spiral slender spirochaetes confirming the diagnosis of primary syphilis [Figure 2]. Treponema pallidum haemagglutination test was also reactive in 1:80 titre; the very low titers of both rapid plasma reagin and Treponema pallidum haemagglutination test possibly indicated the early phase of primary syphilis.

Figure 2:: Smear from ulcer base showing multiple slender spiral spirochaete (in blue circles) (Masson-Fontana silver stain 1000× magnification)

Injection benzathine penicillin 2.4 million units intramuscular (IM) was administered (1.2 million units in each buttock) after a negative penicillin skin sensitivity test. The genital ulcer healed within one month and rapid plasma reagin repeated after three months and became non-reactive.

The mainstay of syphilis diagnosis includes a demonstration of spirochaetes by dark-ground illumination or direct fluorescent antibody microscopy from ulcer exudates or mucocutaneous lesions, or serologic testing for surrogate markers of Treponema pallidum infection.2,3

Direct detection methods are particularly valuable in primary syphilitic chancres, as serological tests may be non-reactive. Among the various direct detection methods, only dark-ground illumination and polymerase chain reaction meet Centers for Disease Control and Prevention (CDC) criteria for laboratory confirmation of syphilis.4

Direct fluorescent antibody is not routinely performed in clinical settings because of high cost and the requirement of expert manpower to accurately interpret the results.

Availability, expertise and operational difficulties associated with dark-ground illumination make it a very subjective test with low sensitivity.

Direct fluorescent antibody, Treponema pallidum specific immunohistochemistry and polymerase chain reaction detection methods are not available at our centre and dark-ground illumination from wet smear failed to demonstrate spirochaetes. Thus, we decided to use Masson-Fontana silver stain for directly staining the spirochaete, to reduce subjective error.

An impression smear was made from the chancre using a sterile glass slide and air-dried. In the laboratory, the smear was first treated by Fontana’s fixative containing formalin and glacial acetic acid. After fixation, it was treated with Fontana’s mordant containing tannic acid to increase the affinity of the stain. Following fixation, the smear was treated with alcohol and covered with Fontana’s stain containing ammoniacal silver nitrate and heated till steam appeared. The smear was washed with water, air-dried and observed using oil immersion microscopy. The silver oxide precipitated on the organism, increasing its diameter and staining it brownish-black.

Further studies are required to ascertain the reliability and validity of this test for diagnosing primary syphilis. Masson-Fontana silver stain provides direct evidence of spirochaetes and hence can be used as a supplemental or confirmatory test for primary syphilis, especially in early cases with ambiguous serology.

Declaration of patient consent

Patient’s consent not required as patients identity is not disclosed or compromised.

Financial support and sponsorship

Nil.

Conflict of interest

There are no conflicts of interest.

References

  1. , . An update on the global epidemiology of syphilis. Curr Epidemiol Rep. 2018;5:24-38.
    [CrossRef] [PubMed] [Google Scholar]
  2. , , , . The immunopathobiology of syphilis: The manifestations and course of syphilis are determined by the level of delayed-type hypersensitivity. Am J Dermatopathol. 2011;33:433-60.
    [CrossRef] [PubMed] [Google Scholar]
  3. , , . Update on syphilis: Resurgence of an old problem. JAMA. 2003;290:1510-4.
    [CrossRef] [PubMed] [Google Scholar]
  4. , , . Sexually acquired syphilis: Laboratory diagnosis, management, and prevention. J Am Acad Dermatol. 2020;82:17-28.
    [CrossRef] [PubMed] [Google Scholar]

Fulltext Views
1,930

PDF downloads
32
View/Download PDF
Download Citations
BibTeX
RIS
Show Sections