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Kala-Azar Diagnosis Techniques Explained

This document summarizes several methods for diagnosing kala-azar (visceral leishmaniasis): Microscopy of peripheral blood, bone marrow, splenic aspirate, or lymph node smears stained with special dyes can reveal the parasite (Leishmania donovani amastigotes) inside macrophages. Culture of tissue or blood on special media can also grow the promastigote form visible under microscopy after 1-4 weeks. Animal inoculation of material into hamsters, while not routine, can demonstrate amastigotes in smears of developing lesions and is very sensitive but takes weeks.

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66 views5 pages

Kala-Azar Diagnosis Techniques Explained

This document summarizes several methods for diagnosing kala-azar (visceral leishmaniasis): Microscopy of peripheral blood, bone marrow, splenic aspirate, or lymph node smears stained with special dyes can reveal the parasite (Leishmania donovani amastigotes) inside macrophages. Culture of tissue or blood on special media can also grow the promastigote form visible under microscopy after 1-4 weeks. Animal inoculation of material into hamsters, while not routine, can demonstrate amastigotes in smears of developing lesions and is very sensitive but takes weeks.

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Megbaru
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We take content rights seriously. If you suspect this is your content, claim it here.
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Diagnosis of kala-azar

• Microscopy
– Sources of specimen
• Peripheral blood
• € Bone marrow
• Splenic aspirate
• Enlarged lymph node
• The smears are stained by Leishman, Giemsa, or Wright’s
stains and examined under oil immersion objective
• Amastigote parasite can be seen within the macrophages,
often in large numbers
• A few extracellular forms can also be seen
Microscopy cont’d
• Peripheral blood smear:
– amastigotes present inside circulating monocytes and less often
in neutrophils
– Sensitivity can be improved by examination of a thick blood film
– also examine buffy coat smear
• show diurnal periodicity
• Bone marrow aspirate:
– sternal marrow (2nd or 3rd intercostal space)
– Iliac crest
• 0.5 mL of marrow fluid
Culture

• Different tissue materials or blood are cultured on NNN


medium
• This is a rabbit blood agar slope consisting of 2 parts of salt
agar and 1 part of defibrinated rabbit blood
• The material is inoculated into the water of condensation and
culture is incubated at 22°–24°C for 1–4 weeks
• At the end of each week, a drop of culture fl uid is examined
for promastigotes under high power objective or phase
contrast illumination
• Other biphasic medium, like Schneider’s drosophila tissue
culture medium with added fetal calf serum can also be used
Animal inoculation

• Animal inoculation is not used for routine diagnosis


• When necessary, Chinese golden hamster is the animal
employed
• The material is inoculated intraperitoneally or n intradermally
into the skin of nose and feet
• The inoculated animals are kept at 23°–26°C
• In positive cases, the amastigote can be demonstrated in
smears taken from ulcers or nodules developing at the sites
of inoculation or from the spleen
• Animal inoculation is a very sensitive method, but takes
several weeks to become positive

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