Diagnostic Techniques in Parasitology

PARASITOLOGY | CHAPTER 2

DETECTION OF PARASITES IN BLOOD THICK BLOOD SMEAR

 Blood protozoa of major clinical significance  Consist of a thick layer of dehemoglobinized
include members of genera: (lysed) red blood cells (RBCs).
 Trypanosoma (T. brucei & T. cruzi)  Most useful for detecting the presence of
 Leishmania (L. donovani, L. tropica, & parasites, because they examine a larger sample
L. braziliensis) of blood (often there are few parasites in the
 Plasmodium (P. falciparum, P. ovale, blood at the time the test is done).
P. malariae, & P. vivax)
 Toxoplasma (T. gondii) THICK BLOOD SMEAR PREPARATION
 Babesia (B. microti) Prepare at least 2 smears per patient
1 Place a small drop of blood in the center of the
THICK AND THIN BLOOD SMEAR pre-cleaned, labelled slide.
ADVANTAGES DISADVANTAGES 2 Using the corner of another slide or an
THICK  the blood  do not permit applicator stick, spread the drop in a circular
BLOOD elements an optimal pattern until it is the size of a dime (1.5 cm2).
SMEAR (Including review of 3 A thick smear of proper density is one which, if
parasites, if parasite placed (wet) over newsprint, allows you to
any) are more morphology barely read the words.
concentrated 4 Lay the slides flat and allow the smears to dry
(app. 30x) thoroughly (protect from dust and insects).
Insufficiently dried smears (and/or smears that
 allow a more are too thick) can detach from the slides during
efficient staining. The risk is increased in smears made
detection of with anticoagulated blood. At room
parasites temperature, drying can take several hours; 30
(increased minutes is the minimum; in the latter case,
sensitivity) handle the smear very delicately during
THIN  Allows for  Lower staining. You can accelerate the drying by
BLOOD optimal review sensitivity in using a fan or hair dryer (use cool setting).
SMEAR of parasite detecting Protect thick smears from hot environments to
morphology parasites due prevent heat-fixing the smear.
 Provides a to dilution of 5 Do not fix thick smears with methanol or heat.
clearer view of blood sample If there will be a delay in staining smears, dip
individual blood the thick smear briefly in water to hemolyse the
elements, RBCs.
facilitating
detailed
analysis

Page 1|2
THIN BLOOD SMEAR so that only WBC, of the infected RBCs
 consist of blood spread in a layer such that platelets and compared to
the thickness decreases progressively toward parasites are uninfected RBCs
the feathered edge. visualized.
 In the feathered edge, the cells should be in a 4 Allow a more Less sensitive than a
monolayer, not touching one another. efficient detection of thick film especially
parasites (increased where there is a low
THIN BLOOD SMEAR PREPARATION sensitivity 11 times parasitemia.
Prepare at least 2 smears per patient than thin smear).
1 Place a small drop of blood on the pre-cleaned, 5 It is not fixed in It is fixed in methanol.
labelled slide, near its frosted end. methanol.
2 Bring another slide at a 30-45° angle up to the 6 Mainly used to Allow the examiner to
drop, allowing the drop to spread along the detect infection and identify malaria
contact line of the 2 slides. to estimate species, quantify
3 Quickly push the upper (spreader) slide toward parasitemia. parasitemia, and
the unfrosted end of the lower slide. recognize parasite
4 Make sure that the smears have a good forms like schizonts
feathered edge. This is achieved by using the and gametocytes.
correct amount of blood and spreading
technique. Sedimentation Concentration Technique
5 Allow the thin smears to dry. (They dry much  Use solutions of lower specific gravity than the
faster than the thick smears, and are less parasitic organisms, thus concentrating the
subject to detachment because they will be latter in the sediment.
fixed.)  It can be used for separation of protozoan
6 Fix the smears by dipping them in absolute cysts or oocysts and helminth eggs and
methanol. larvae from stool samples.

DIFFERENCES BETWEEN THICK & THIN BLOOD

SMEARS

THICK THIN
1 most useful for Helps to discover which
detecting the species of parasite is
presence of causing the infection.
parasites
2 A drop of blood on a A drop of blood that is
glass slide. spread across a large
area of the slide.
3 The blood films must The purpose is to allow
be laked before or malarial parasites to
during staining to be seen within the RBC
rupture all the RBC and to assess the size

Page 2|2