How to do

a skin smear examination

for leprosy
dr.Lily Pertiwi Kalalo,SpPK
Why take a skin smear?

To confirm a diagnosis of skin smear-

positive multibacillary leprosy in a suspect.

To help diagnose multibacillary relapse in

a patient who has previously been treated.

To help with the classification of new

patients.
How to do a skin smear examination for leprosy
Explain to the patient
sk the patient to sit down and
relax. !xplain what
you want to do and why it is
necessary. nswer any
"uestions. #btain the patient$s
permission to proceed
and enter the details on the
re"uest form.
How to do a skin smear examination for leprosy

Remember: Taking a skin smear is an

invasive procedure. Wash your hands%
wear gloves% and

&se sterilised e"uipment and a new blade

for each patient. 'o not take skin smears
unnecessarily.
How to do a skin smear examination for leprosy
(reparing to take a skin smear
Equipments
How to do a skin smear examination for leprosy
Gloves
Spirit lamp



Scalpel
handle
and new
blades
Dressing strips Alkohol Swab
Slide box and new slides
Select the sites
Take a smear from two sites only:
1. One ear lobe. 2. One lesion (most active-
looking lesion, but not on the face).
!ctive means raise" an" re""ish in colour.
How to do a skin smear examination for leprosy
# $f there is no suitable skin lesion, take
the secon" smear from the other ear
lobe, or from a site where active lesions
were
# originally recor"e" or where a %revious
smear was %ositive.
How to do a skin smear examination for leprosy
&any %rogrammes tra"itionally took smears from
four or even si' sites,
but two sites are now consi"ere" a"e(uate
in most cases.
How to take a skin smear
How to do a skin smear examination for leprosy

Wash your hands

and put on gloves.
How to do a skin smear examination for leprosy
Take a new% clean% unscratched microscope
slide. &sing a slide marker% write the patient
identification )*'+ number at the bottom of
the slide
This number must be on the
re"uest form.
How to do a skin smear examination for leprosy
#Clean the skin at the smear sites with a
cotton wad drenched in alcohol.Let it
dry.
Put a new blade on the scalpel handle.
Pinch the skin firmly between your
thumb and forefiner! press out the
blood.
How to do a skin smear examination for leprosy
)kin incision about * mm long an" 2 mm "ee%
#+ee% on %inching to make sure the cut remains
bloo"less. $f blee"ing, wi%e the bloo" away with cotton
wa".
# Turn the scal%el ,-. an" hol" it at a right angle to
the cut.
#)cra%e insi"e the cut once or twice with the si"e of
the scal%el,to collect tissue flui" an" %ul%. There
shoul" be no bloo" in the s%ecimen
# )to% %inching the skin an" absorb any blee"ing with a
wa" of cotton.
How to do a skin smear examination for leprosy
# Spread the material scraped from the
incision onto the slide,
# on the same side as the "# number.
Spread it e$enly with the
# flat of the scalpel, makin a circle % mm
in diameter
# /ub the scal%el with a cotton wa"
"renche" in alcohol.
# 0ass the bla"e through the flame of the
s%irit burner for 1 to 2 secon"s.
# 3et it cool without touching anything.
# /e%eat the ste%s above for the secon"
site.
# )%rea" this smear ne't to, but not
touching, the first one.
# 4iscar" the scal%el bla"e safely.
# 4ress the woun"s an" thank the %atient.
#3et the sli"e "ry for 1* min at room tem%erature.
# 5i' the smears by %assing the sli"e, with the
smears u%war"s, slowly through the flame of a
s%irit burner, 1 times.
How to do a skin smear examination for leprosy
How to stain a skin smear

,tain the smears using the hot -iehl-.eelsen

method. ,tain with /0 carbol fuchsin% which
colours everything red.

Wash out the stain with /0 acid-alcohol%

which removes the stain from everything
except M. leprae.

1ounter-stain the slide with 2.30 methylene

blue.

The leprosy bacilli will be

seen as red rods on a blue background.

How to do a skin smear examination for leprosy
Equipment
How to do a skin smear examination for leprosy
1% carbol fuchsin solution
1% acidalcohol
!."% meth#lene blue solution
Staining
How to do a skin smear examination for leprosy
# filter the 16 carbol fuchsin.
# 7over the whole sli"e with 16 carbol fuchsin
solution (&).
# 8eat the sli"e gently by hol"ing a burning s%irit
lam% un"erneath it until va%our begins to rise from
the carbol fuchsin ('). /e%eat this 1 times "uring
a %erio" of * minutes. &ake sure the stain "oes
not boil. 9ash gently un"er a running ta% (().
# /inse until the run-off water is colourles.
)1+.
)2+.
)3+.
Decolourising
How to do a skin smear examination for leprosy
7over with 16
aci"-alcohol
for 1- secon"s
)5+.
/inse gently with water
)4+.
Counter-staining
How to do a skin smear examination for leprosy
5inse with water )7+
let the slide dry in the drying rack in an
inclined position%with the smeared side
downwards )+.
The slide is now ready to be read.
1over with 2.30 methylene
blue
for /minute )!+.
How to read a skin smear
How to do a skin smear examination for leprosy
:ou nee" a microsco%e with a
1-' eye %iece an" 1-' an" 1--'
ob;ectives.
)tart the e'amination using the
1-' ob;ective.
immersion oil
)oli" bacilli may
suggest the %resence of viable organisms an" may be seen in
new, untreate" cases or in rela%se cases.
How to do a skin smear examination for leprosy
soli"s
<lobi fragmente"
granuler
='amine a%%ro'imately 1-- fiel"s %er smear.
>acteriological $n"e' (>$)
- - bacilli in 1-- fiel"s
1? 1 @ 1- bacilli in 1-- fiel"s
2? 1 @ 1- bacilli in 1- fiel"s
1? 1 @ 1- bacilli, on average, in each fiel"
2? 1- @ 1-- bacilli, on average, in each fiel"
*? 1-- @ 1--- bacilli, on average, in each fiel"
A? B 1--- bacilli, on average, in each fiel"
.ote6 5eport the 7* for both smears on the slide. 8or smear-positive patients% either
the average 7* or the highest 7* will be taken as the 7* for that patient.
"# $"or%ological #n&ex'
(erbandingan 9umlah bakteri tahan
asam yang utuh )solid+ berwarna
merah dengan total bakteri tahan
asam.

No Lokasi Pengambilan Sampel Kepadatan
Ridley
Solid
/ utuh
Fragmented
/ granulated
1. Cuping telinga kanan + 5 30 70
2. Cuping telinga kiri + 5 25 75
3. Lengan kanan + 4 20 80
4. Lengan kiri + 4 26 74
5. Bercak tersangka + 5 35 65
Jumlah +23 136 364
Contoh perhitungan (# &an "#
(eriksa kepadatan 5idley sediaan tiap lokasi
Hitung /22 7T dari sediaan tiap lokasi dan catat hasilny
7acteriological *ndex : 3; 6 < : 4%=
/;=
>orfological *ndex : -------------- ? /220 : 3@%30 atau /;= 6 < :
3@%30
/;=A;=4