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PARASITOLOGY
2.1B DIAGNOSTIC PARASITOLOGY

DIAGNOSTIC PARASITOLOGY
GENERAL PRINCIPLES
1. Consider the possibility of parasitic infection in diagnosis. Think of it.
2. Take a complete history of travel, includes the recent and past. Where
have you been?
3. Should have knowledge of the parasite biology.
4. Consider the facilities in deciding what lab tests are necessary.
5. Finally, interpret the result of the tests in the light of the clinical picture,
then treat the disease not the tests

METHODS OF DIAGNOSIS OF PARASITIC INFECTIONS

Demonstration or Identification of parasites
developmental stages

eggs, larvae and adults

cysts, oocysts and trophozoites

trophozoites, schizonts and gametocytes

SPECIMENS FOR PARASITIC EXAMINATION

Stool
Blood
Sputum
Urine and Genital Secretions
Tissue aspirate and tissue biopsy material
Eye Specimens
Cerebrospinal Fluid (CSF)
Mouth Scrapings and Nasal Discharge
Skin Snips

FACTORS TO CONSIDER IN COLLECTION & PROCESSING OF

STOOL SAMPLES
Intake of drugs/substances
Intake of anti-malarial/antibiotics
Collect in tight fitting lid container

Contamination w/ urine, water and soil

Amount of sample

Label
Time frame (age of stool sample)
Preservation

Preservatives

Temporary storage

Never freeze or keep the sample in an incubator

COLLECTION: FIXATIVES
Formalin
PVA (Mercuric oxide)
Sodium Acetate formalin
Modified PVA (copper and zinc sulfate)
Alternative Single-Vial systems (non-toxic fixatives)

PROCESSING OF STOOL SAMPLE

Method of examination

Macroscopic or Physical

Consistency

Color

Microscopic

Stages of parasites
MACROSCOPIC

Detection of host immune response to the parasites

antigen
and antibody
different immunodiagnostic tests

THINGS TO CONSIDER IN DIAGNOSTIC PARASITOLOGY

Proper collection and transport of sample
Processing of specimen prior to examination
Skills of the laboratory personnel
Quality of equipments

CONSISTENCY
Hard
Soft
Mushy
Loose
Diarrheic
Watery, liquid
Formed
Semiformed

COLOR
Dark brown
Black
Brown
Pale brown
Clay
Yellow
Red-brown
Green, others

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Parasitology

2.1B DIAGNOSTIC PARASITOLOGY

CONSISTENCY

DISTRIBUTION OF PROTOZOA IN RELATION TO CONSISTENCY

CONCENTRATION TECHNIQUES
SEDIMENTATION
Formalin Ethyl Acetate
Provides good recovery
Easy to perform
Dis. - contains more fecal
debris

MICROSCOPIC
Different stages of the parasites
Elements (WBC,RBC, macrophages and others)
Normal fecal constituents like fat globules, fibres and others

MICROSCOPIC TECHNIQUES
Direct fecal smear (DFS)
Concentration techniques
Permanent stain
Culture method
Egg counting
Cellophane Tape preparation

DIRECT WET PREPARATION

UNSTAINED
STAINED
NSS
Lugol's solution
Use to detect motile
Enhance the details of
protozoan parasite
protozoan cysts
Dis. - low yield of
Dis. - Iodine kills
parasites
trophozoites

FLOTATION
Zinc sulfate
Yields cleaner preparation
Easy to examine
Dis. - some parasites will
be missed

PERMANENT STAINS
TRICHROME
IRON HEMATOXYLIN
Widely used

Time consuming
Long shelf life

Reveals excellent
Easy to perform
morphology of the
Differentiates the color of
intestinal protozoans
parasites and background

Nuclear details are

stained clearer and
sharper than trichrome

OTHER PROCEDURES
STOOL CULTURE METHOD
EGG COUNTING
Harada-Mori or Test
Kato Katz
tube culture method
Kato Thick smear

OTHER INTESTINAL SPECIMENS

Duodenal materials
Sigmoidoscopy materials
Sample from Cellophane Tape preparation

DUODENAL MATERIALS

Collected by nasogastric
intubation

Or enteric capsule test

(Enterotest)

Giardia intestinalis trop,

I. Belli, Cryptosporidium
spp, S. stercoralis, and
eggs of F. hepatica or C.
sinensis

SIGMOIDOSCOPY MATERIALS

Helpful for detecting

E.histolytica

Obtained by aspiration or
scraping fr ulcers

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Parasitology

2.1B DIAGNOSTIC PARASITOLOGY

CELLOPHANE TAPE

S. stercolaris hyperinfection
E. histolytica and E. gingivalis
A. lumbrecoides and hookworm
Examine directly using wet preps and permanent
stain
URINE & GENITAL SECRETIONS
S. haematobium eggs
Microfilariae fr patients w/ heavy infection
Sample should be centrifuged and examine the
sediment
T. vaginalis troph (urine and genital secretions)
Genital secretions are collected in a swab or in a
cup, examined by wet preps to demonstrate motile
trophozoite

EYE SPECIMENS
CORNEAL SCRAPING FOR A. KERATITIS

Culture

Stained with calcofluor using fluorescent

microscope

Histologic method

Loa loa and T. gondii

MOUTH SCRAPINGS AND NASAL DISCHARGE

Mouth scrapings for detection of E. gingivalis and T.

tenax

Recovery of N. fowleri fr nasal samples

Both are collected on a swab or in a cup

Wet preps

BLOOD
COLLECTION
Aseptic technique
whole blood fr fingertip or earlobe
or venipuncture specimen w/ anticoagulant (EDTA)
Time of collection
L. donovani, Trypanosoma spp., Plasmodium and
Babesia specie

PROCESSING
Thick and thin smear
Permanent stains
Knott method
Buffy coat slides
Culture using NNN medium

OTHER LAB METHODS

CULTURE
ANIMAL INOCULATION
E. histolytica

Specimens fr patients
T. vaginalis
suspected of suffering fr
Leishamnia spp.
Leismania,
T. cruzi
Trypanosoma and
T. gondii
Toxoplasma

Collected aseptically

Mice, guinea pigs and

hamsters
XENODIAGNOSIS

Chaga's disease

T. cruzi

CSF AND OTHER STERILE FLUID

Collect aseptically
Examined promptly
Wet preparation and/or permanent stains
N. fowleri, Acanthamoeba spp, Trypanosoma spp,T. Gondii, T.
solium cysticercus larvae and Echinococcus spp
Culture on non-nutrient agar if Naeglaria and Acanthamoeba
are the suspected pathogens

TISSUE & BIOPSY SPECIMENS

Recommended for the recovery of many parasites
includes intracellular organisms
Surgical removal of the specimens for Leishmania
spp and T. gondii
Liver abscess for E. histolytica
Trypanosoma spp and T. spiralis
SPUTUM
Collected and tested from patients suspected of
being infected by the fluke. P. westermani

SKIN SNIPS
ONCHOCERCA VOLVOLUS
Two techniques of collection:
Scleral punch into skin
Using razor blade with w/c small cut into skin is
made
Put sample into saline solution and incubate for
30min
Examine sample and look for the jerky movement of
the microfilariae

RECENT ADVANCES IN DIAGNOSTIC PARASITOLOGY

MICROSCOPY
IMMUNODIAGNOSIS

Rapid diagnostic test

MOLECULAR DIAGNOSIS

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Parasitology

2.1B DIAGNOSTIC PARASITOLOGY

MICROSCOPY
FLUORESCENT

Malaria, filariasis, leishmaniasis,

trypanosomiasis, and onchocerciasis for
both method
Toxoplasmosis, taeniasis and
trichomoniasis for PCR only
END OF TRANS

Trophozoites of P.
falciparum
stained with AO in the
QBC
UV fluorescence method.

Trophozoites of P.
falciparum stained with
BCP in the fluorescence
method.
IMMUNODIAGNOSIS
Immunofluorescent Assay
Enzyme Linked Immunosorbent Assay
Indirect Hemagglutination Assay
Radioimmunoassay
Dot blot
RAPID DIAGNOSTIC TESTS
RDTS FOR MALARIA
RDTS FOR MALARIA
HRP based assay
pLDH based assay

RDTS FOR OTHER PARASITES

FAST ( fast agglutinating screening test) for visceral leishmania
LAT (latex agglutination test) for Taenia solium
Immunochromatographic test for filaria
MOLECULAR DIAGNOSIS
DNA-BASED METHOD

Detection of parasite using DNA-probe

Hybridization assays

Detection of specific nucleic acid

sequences

PCR- based assays

Amebiasis, used to differentiate

E. histolytica and E.dispar
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