CLINICAL PARASITOLOGY

Bachelor of Science in Medical Laboratory Science Notes 2. Concentration Techniques

2nd Semester, A.Y 2019-2020
Compiled by MIGUEL LORENZ C. PARAWAN
*Source/s are varied.
PROCEDURES COMMONLY PERFORMED IN
CLINICAL PARASITOLOGY
- separates parasites from fecal debris
- increase the chances of detecting parasitic
organisms when these are small in numbers
[Flotation Technique and Sedimentation
Technique]

1. Direct Fecal Smear (DFS) Flotation Technique

- most useful for the diagnosis of protozoal  Most frequently used: Zinc Sulfate
parasites which have motile trophozoite (ZnSO4) or Sheather’s sugar (454g
stages that are passed in the feces granulated sugar + 355mL tap
- should be performed on concentrated water + 6mL full-strength [37%]
specimens formaldehyde)
- specimens used should be a fresh feces  Uses solutions which have higher
- smear should be thin enough so that specific gravity than the
printed page can be read through it organisms to be floated so that
LOW POWER: scans for large helminth organisms rise to the top and the
eggs or larvae debris sinks to the bottom
HIGH POWER: detects and identifies
smaller parasites and ADVANTAGE: produces a cleaner
larger helminth eggs or material than the
larvae sedimentation technique
- parasites detected are reported out by DISADVANTAGE: the walls of eggs and
their scientific name and quantity cysts will often
observed collapse, thus
hindering identification
INDICATION: Motile protozoan ; some parasites don’t
s

trophozoites (feces must be body temp.) float

LIMITATION: Small smaple size Sedimentation Technique

[Saline and Iodine Wet Mount
Preparations]
- evaluation of saline direct mounts:
 provides opportunity to study the
motility of any trophozoites
present.
- addition of iodine:
 stains the internal structures
necessary for identification of the
cysts of some amoeba and other
protozoa
 Trophozoites are rapidly killed and
are sometimes unidentifiable
upon addition of iodine.
Video Reference:
https://youtube.com/watch?v=dklHE5gJ3
94
 Uses solutions of lower specific
gravity than the parasitic
organisms, thus concentrating the
latter in the sediment
 Recommended for general
diagnostic laboratories because
they are easier to perform and
less prone to technical errors
 CDC sedimentation technique:
Formalin-ethyl acetate technique
- a diphasic sedimentation
technique that avoids the
problems of flammability of ether,
and which can be used with
specimens preserved in formalin,
MIF or SAF.
Video References:
https://youtube.com/watch?v=qLnpo5dveV4
https://youtube.com/watch?v=GTJzxZRo49A
3. Kato-Katz Technique
- used for qualitative and
semi-quantitative diagnosis of intestinal
helminthic infestations
- needs thick smear
- typically diagnoses Ascaris lumbricoides,
Trichuris trichiura, hookworm, and
especially Schistosoma spp.
- WHO recommends this technique in areas
with moderate to high transmission rates of
soil transmitted helminths or intestinal
schistosomiasis
- people infected with STH or intestinal
schistosomes pass the eggs of the worms
through their feces
PRINCIPLE:
1. Feces are pressed through a mesh
screen to remove large particles
2. After filling the hole, the template is
removed and the remaining sample is
covered with a piece of cellophane soaked
in glycerol.
Note: Glycerol clears the fecal material
from around the eggs.
3. The eggs are then counted, and the
number calculated per gram of feces.
Video Reference:
https://youtube.com/watch?v=WpcZejHa_jM
4. Harada Mori Culture
- method of incubating fecal material on a
filter paper strip in a test tube containing
water
- used to culture and recover nematode
larvae (Strongyloides stercoralis,
hookworm)
PRINCIPLE:
 Employs a filter paper to which fecal
material is added and a test tube to
which filter paper is inserted.
 Moisture is provided by adding water
to the tube.
 Water continuously soaks the filter
paper by capillary action.
 Incubation under suitable conditions
facors hatching of ova or the
development of larvae.
 Fecal cultures are to be cultured and
not refrigerated, since some parasites
(especially Necator americanus) are
susceptible to cold and may fail to
develop after refrigeration.
Video Reference:
https://youtube.com/watch?v=Rcw4irQpNlc
5. Scotch Tape Method
- collection and examination of a sample
from the skin around the anus
- used to identify pinworms and pinworm
eggs
PRINCIPLE:
 Female pinworm (Enterobius
vermicularis) migrates out of the anus
at night to deposit her eggs on the
perianal region.
 These eggs are easily recovered by
pressing a piece of scotch tape against
the perianal region.
 The tape is then placed on a
microscope slide and submitted to the
laboratory for examination.
 The specimen should be obtained first
thing in the morning before the patient
bathes of defecates.
Video Reference:
https://youtube.com/watch?v=1KMvfkbjHng
6. Blood Smear
- used to look for specific parasites that are
found in the blood
- very time consuming
- parasitic diseases such as filariasis,
malaria, or babesiosis can be diagnosed
Note: Parasites in the peripheral blood may
be present in very small numbers. Careful
examination of the blood smear is required
before reporting the smear out as “no
parasites seen.”
[Thin Blood Smear and Thick Blood Smear]
Thin Blood Smear
 Used for the specific identification
of malarial parasites,
trypanosomes, and microfilariae
 Made in the same way smears are
made for hematological study
ADVANTAGE: preserves the structure
of the parasites with a
minimum distortion
Thick Blood Smear
 Preferred over thin film for
diagnosis of malaria because it
reveals comparatively quicky
sparse or scanty infections such
as those occuring in new or
chronic cases
 Enables one to examine 10-50
times as much blood as the thin
films in the same length of time.
PRINCIPLE:
 This test is done by placing a drop of
blood on a microscope slide.
 The slide is then stained and examined
under a microscope.
Video References:
https://youtube.com/watch?v=acoALifVvb8
https://youtube.com/watch?v=WPP7AjmStBg
8. Occult Blood Test
- may indicate lesions in the intestinal tract
due to parasitic infections, cancerous
tumors, bleeding ulcers, and other disease
processes
- stools with large amount of occult blood
are black and have a tarry consistency
CHEMICAL REACTION:
1. Hemoglobin + Developer
Hb + 2 H2O2 = 2 H2O + O2
2. Oxidation of Guaiac
O2 + Guaiac = Oxidized Guaiac
[colorless] [blue]
Note:
 Presence of blue color upon
completion of the test indicates a
positive response.
 A positive and negative control must
be run concurrently with the patient
sample.
 It is recommended that the patient be
placed on a high residue diet starting 2
days before and continuing through
the test period.
Video Reference:
https://youtube.com/watch?v=z8d1LT0VnXI

7. Circumoval Precipitin (COPT)

- used to detect serum antibodies to
Schistosoma mansoni, Schistosoma
haematobium, or both species by using
eggs of either species of schistosome.
- complicated and very time consuming
(~48 hrs)
- requires microscopy

PRINCIPLE:
 Eggs of either the two Schistosoma spp.
Were aderquate as antigens but in
general, the sera of S.
mansoni-infected individuals caused
more circumoval precipitates for form
around S. mansoni eggs (indicating
higher sensitivity) than did the sera of
S. haematobium-infected individuals.