Lab Diagnosis Of Leprosy

Moderator : Dr. Bhawesh koirala

Resource: Mr. Tejendra Pandit
Speaker: MSM
 OBJECTIVES:
•TO DESCRIBE THE LABORATORY
DIAGONOSIS OF MYCOBACTERIUM LEPRAE
 BRIEF INTRODUCTION:
•It is primarily a granulomatous disease of
the peripheral nerves and mucosa of the
upper respiratory tract; skin lesions are the
primary external sign.
•Caused by the bacteria Mycobacterium leprae
and Mycobacterium lepromatosis.
 LAB DIAGNOSIS
•Sample collection
•Microscopy
•Culture
•Serology
•Molecular method
 SAMPLE COLLECTION
• Specimen are collected from
A. Nasal mucosa,
B. Skin lesion
C.Both ear lobules
D.Right elbow
E Middle phalanx of left finger
F. Buttocks
• Skin is pinched and cut about 5mm and a
deep infiltrated layer is
taken with a scalpel. – SKIN LESION
 NASAL MUCOSA
Blunt, narrow scalpel is
introduced into the
nose and a piece of
nasal mucous
membrane is taken
NERVE BIOPSY:
Nerve biopsy: Sample is
collected from thickened
nerves(eg sural nerve)
and submitted for
histopathological
examination.
 MICROSCOPY
•Microscopy: acid fast
bacilli stained with ZN
staining

•Ziehl-Neelsen technique
use in this uses 5%
instead of 20% sulphuric
acid for decolourisation.
 GRADING:
•1-10 bacilli in 100 fields =1+
•1-10 bacilli in 10 fields =2+
•1-10 bacilli per fields =3+
•10-100 bacilli per fields =4+
•100-1000 bacilli per fields =5+
•More than 1000 bacilli per fields =6+
 BACTERIOLOGICAL INDEX
•Number of total bacilli in a tissue.

• B.I = Total score in smear

No. of smear
for e.g 16/8 so,B.I 2
• Minimum of 4 skin lesions,a nasal swab
& both the ear lobes are to be examined.
 MORPHOLOGICAL
INDEX
• Morphological index(M.I.) is expressed as
the percentage of solid fragmented
granular bacilli (SFGB)or uniformly stained
bacilli out of the total number of bacilli
counted
• For assessing the progress of
patients on chemotherapy.
 CULTURE
ANIMAL INOCULATION

Mouse foot pad inoculation has been reported

to be more sensitive than skin slit smears.
Disadvantage
It is unsuitable for routine diagnosis and
feasible only for drug potency or resistance
testing research studies.
• Armadillo: the nine banded armadillo is
highly susceptible to infection with M.leprae.
 Serology:
Serology: Detection of antibody against
M.leprae phenolic glycolipid antigen has been
claimed to specific diagnostic test.

Their role has not yet been accepted for

diagnosis.
 MOLECULAR METHODS:
Molecular methods:
Attempts of it’s development are in progress.
 LEPROMIN TEST
• Delayed type of hypersensitive reaction.
• First described by Mitsuda in 1919.
• Lepromins used as antigens may be of
human origin (lepromin H) or armadillo
derived (lepromin A).
PROCEDURE :
Carried out by the intradermal
injection of 0.1 ml of lepromin.
BIPHASIC RESPONSE:
1) Early reaction of Fernandez
2) Late reaction of Mitsuda
USES OF LEPROMIN TEST:
a) Classification ofleprosy:
- Positive in tuberculoid leprosy
- Negative in lepromatous leprosy
b) Assessment of prognosis:
- Positive lepromin test indicates a good
prognosis.
- Negative lepromin test indicates a bad
prognosis.
c) Assessment of resistance:
- To assess the resistance of an individual to
leprosy.
- Resistance is indicated by positive
lepromin test.
 OTHER TEST
• Polymerase chain reaction (PCR):

• Lymphocyte migration inhibition test (LMIT):

 REFERENCES :
•Textbook of microbiology by P.Chakarborty.

•Jawetz’s Review of medical microbiology

•Harrison’s principles of internal medicine 18

edition ,page:1363-1364