Pharmaceutics – level 3 &4 –parasitology – LAB 3 T/ Mohammed Hail Al Adeemy

Diagnosis of E.histolytica
E. histolytica transmitted by the faecal-oral route with
infective cysts being ingested in food, water, or from
hands contaminated with faeces.
E. histolytica has a direct life cycle as shown in Figure :

Laboratory Diagnosis of Intestinal Amoebiasis :

Sample collection :
Stool is the specimen of choice. Minimum of three stool
samples should be collected on consecutive days as
amoebae are shed intermittently.
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Pharmaceutics – level 3 &4 –parasitology – LAB 3 T/ Mohammed Hail Al Adeemy

Stool specimen should be collected in wide mouthed

clean container. Other samples include rectal exudates
and rectal ulcer tissues collected by colonoscopy.
It should be examined immediately within 1–2 hours of
collection (watery 15 minutes) or can be preserved in
polyvinyl alcohol or formalin.

Stool microscopy :
Direct examination of stool by saline and iodine mount
is done to demonstrate: trophozoites and cysts as
follow :
1- Place a drop of saline on the one end of the slide.
Using a wire loop or piece of stick, place a small
amount of specimen on saline and mix it. cover
with a cover glass and using a tissue, press gently
on the cover glass to make a thin preparation.
2- Place a drop of iodine on the other end of the slide.
Mix a small amount of the specimen with the
iodine and cover with a cover glass.

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Pharmaceutics – level 3 &4 –parasitology – LAB 3 T/ Mohammed Hail Al Adeemy

3- Examine immediately the preparations

microscopically, first using the 10X objective with
the condenser iris closed sufficiently to give good
contrast. Use the 40X objective to identify motile E.
histolytica trophozoites or cysts.

NOTE :
The laboratory diagnosis of amoebic dysentery is by
finding E. histolytica trophozoites in a fresh dysenteric
faecal specimen or rectal scrape. Specimens must be
examined without delay, otherwise identification of the
trophozoites becomes impossible because the amoebae
lose their motility, extrude food vacuoles containing red
cells, and round up.

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Pharmaceutics – level 3 &4 –parasitology – LAB 3 T/ Mohammed Hail Al Adeemy

Stool culture :
Polyxenic and axenic culture.

Stool antigen detection :

(copro-antigen) CIEP, ELISA, ICT.

Serology :
- Amoebic antigen—ELISA.
- Amoeboic antibody—IHA, ELISA and IFA.

Molecular diagnosis :
Nested multiplex PCR and real time PCR.

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