SLIT SKIN SMEAR

Muhamed Raeez P
CONTENTS
◦ Introduction
◦ Principle
◦ Method
◦ Staining of skin smears
◦ Microscopic Examination of skin smears
◦ Interpretation
INTRODUCTION
◦ Slit skin smear is a rapid, valuable and cost-effective method to diagnose
Hansen’s disease.
◦ Estimates the density of acid-fast bacteria.
◦ Reported as Bacterial index (BI) and Morphological index (MI).
◦ Determines the type and severity of disease as well as assessment of treatment
response and prognosis.
PRINCIPLE
◦ Serous fluid and dermal tissue from the skin is smeared and stained.
◦ It demonstrates Mycobacterium leprae.
◦ Smears are usually taken from five sites- both ear lobules, lateral parts of
forehead, near both eyebrows and active and suspicious lesions.
◦ Smear from dorsa of finger is earliest indication of impending relapse.
◦ Depending on number and morphology of bacilli, leprosy is classified
 METHODS

◦ Universal precautions should be observed in obtaining skin smears.

◦ The skin cleansed with 70% alcohol and air dried or wiped dry with cotton.
◦ Skin is pressed with index and thumb to make area relatively avascular.
◦ A 3-5 mm long and 2-3 mm deep incision is made with 15 number Bard-Parker
blade.
◦ The inner surface of the wound is scraped with the blade held at a right angle to
the incision.
◦ Upon scraping, tissue fluid and dermal tissue are obtained.
◦ The material is transferred to clean microscope slide, a moderately thick smear
with a visible uniform opacity is made.
◦ The smear is made in a circular manner (5-7mm) beginning peripherally and
ending in the centre, leaving a central “button” (2-4 mm)
STAINING OF SKIN SMEAR
◦ The slide is air dried or gently heated between 40 and 50 degree Celsius.
◦ Place slides on a staining rack and flood the slide with concentrated Ziehl-Neelson
or carbol-fuchsin for 20 minutes.
◦ Gently rinse slides well with tap water to remove excess stain.
◦ Decolorize with 5% sulphuric acid (H2SO4) in 70% ethanol or 5% H2SO4 for 1
minute.
◦ Gently rinse slides thoroughly with tap water.
◦ Counterstain with alkaline methylene blue for 30 sec to 1 min.
◦ Gently rinse well with tap water and air dry.
MICROSCOPIC EXAMINATION OF
SKIN SMEARS
◦ The stained smears are examined under microscope using oil
immersion objective (100x) to determine total no. of bacilli.
◦ Rod-shaped pink colored bacilli lying singly or in groups are
demonstrated on a blue background.
 INTERPRETATION
◦ Smears should be looked for both live and dead bacilli. Two
indices are used BI and MI
◦ Bacteriological index counts both viable and dead bacilli.
◦ Morphological index calculates only viable bacilli
◦ Live bacilli are uniformly stained rods having rounded ends,
parallel sides and length about 4 times width
◦ Dead bacilli are seen as irregularly stained (fragmented) rods.
◦ The results are reported on 0 to 6+ semi-logarithmic scale
using numerical code. This is an indicator of the total
bacillary load of the patient.
◦ Morphological index estimates the percentage of solid
staining bacilli lying singly, after examining 200 red staining
bacilli.
◦ Helps to ascertain whether the patient is responding to
treatment or the disease is still active.
INTERPRETATION
Grading of Bacteriological Index (BI)

Grades (BI) DESCRIPTION

1+ 1-10 bacilli per 100 fields.
2+ 1-10 bacilli per 10 fields.
3+ 1-10 bacilli per oil immersion
field.
4+ 10-100 bacilli per oil immersion
field.
5+ 100-1,000 bacilli per oil
immersion field.
6+ Over 1,000 bacilli per oil
immersion field.
REFERENCES
◦ IADVL’s CONCISE TEXTBOOK OF DERMATOLOGY 2nd EDITION.
◦ GOOGLE IMAGES.
THANK YOU