DIAGNOSTIC PARASITOLOGY:

PROCEDURES AND
PRACTICES

Myra S. Mistica, MPH

University Researcher
Department of Parasitology
College of Public Health
University of the Philippines Manila
 Course Objectives

At the end of the training course

 identify important morphologic

features of parasites and
medically important arthropods

 perform various laboratory

techniques in the diagnosis of
parasitic infections
 Diagnostic Parasitology

• very important role in

establishing parasitic infections

• correct parasite identification

aids in selection of appropriate
drugs for treatment
Specimens for Diagnosis of
Parasitic Infections
• Feces, stool sample - STH, food
and waterborne parasites,
protozoans
• Blood/ serum – malaria, filaria,
schistosomiasis
• Sputum - paragonimiasis,
strongyloidiasis
• Urine - schistosomiasis
• Perianal swab – enterobiasis,
tapeworm
Collection of Fecal Specimen

• Clean, wide-mouth container

Considerations
• Urine in feces – should be
avoided
• Collect feces – not from soil
• Appropriate amount – size of
one’s thumb
- formed
- semi formed
- watery
• How many stool examinations?
– 3 times
 Collection of Blood
Specimen
• Through finger prick or
venipuncture
• Type of container – depends on
examination/test to be performed
• Malaria examination - thin and
thick smear
• Microfilaria examination - thick
blood film
• Serological tests - vial or tube
without anticoagulant
Blood sample preparation for microscopic examination

Thin blood
film

Thin blood film

Thick
blood
film Thick blood film
9
Collection of Sputum
Specimen
• Early morning after waking up
• Clean sputum cups
• Patients are advised to gargle
first before collection
• Breathe deeply before
expectorating
 Techniques
• Wet Smear Method
– DFS

• Stool Concentration Technique

– FECT – Formalin Ether
Concentration Technique
– MIFC – Merthiolate Iodine
Formaldehyde Concentration
Technique
– AECT – Acid Ether Concentration
Technique
– Kato Katz
 Techniques

• Staining techniques
– Kinyoun’s staining (modified
acid-fast)

• Other techniques
– Scotch Tape Swab
– Tapeworm segment
examination
 Techniques
• Blood/serum
– Thick blood smear
– Thick and thin blood smear
– Knott’s concentration
– COPT
• Sputum
– 3% NaOH Concentration
Technique
• Perianal swab
– Scotch tape swab
 Parasitologic Methods

Qualitative
Clinical practice/routine
examination
Direct fecal smear
Kato thick method

Quantitative
Epidemiological studies
Control program
Clinical trials
Kato Katz method
Direct fecal smear
- easiest laboratory procedure
- simple, cheap, quick procedure

Kato Katz technique

- more commonly used for
epidemiologic studies, control
programs and clinical trials
- processed specimen can be kept
for 2 weeks

Kato thick
DIRECT FECAL SMEAR (DFS)

Examines ~ 2 mg of feces

Ideal for diarrheic and

soft stool specimens
-----------------------------------------------

Examines ~ 40-50 mg of feces

Ideal for intestinal helminths

and non-diarrheic stool specimens

KATO KATZ METHOD or

MODIFIED KATO THICK METHOD
Kato Katz – qualitative and
quantitative , uses a template

All eggs are counted by species.

The absolute count of eggs per
species is multiplied by a factor
(23, 24) to come up with number
of eggs/ gram feces.

Kato thick – qualitative, no

template
 WORM BURDEN -
WHO CLASSIFICATION
LIGHT MODERATE HEAVY
ASCARIS 1-4999 5000-49999 > 50000

TRICHURIS 1-999 1000 – 9999 > 10000

HOOKWORM 1- 1999 2000- 3999 > 4000

SCHISTOSOMA 1- 99 100 – 399 > 400

 Artifacts

• Air bubbles
• Oil droplets
• Macrophages/PMNs
• Starch granules
• Fungal spores/pollen
grains
• Fibers – meat or plant
origin
Artifacts
Oil droplets
Air bubbles
Plant fibers
Starch granules
Fungal spores/pollen
grains/algal spores
Charcot Leyden Crystals
Mite egg
 Diagnostic Parasitology
Laboratory
• DFS/Kato thick
• Kato Katz
• FECT
• Kinyoun’s staining
• COPT
• Blood smear examination
• IFAT amoeba/malaria
• Identification of
adults/segments
 Medical Entomology
Laboratory
• Bioefficacy evaluations of
products against
mosquitoes, cockroaches,
flies
• Identification of medically
important arthropods
 References

• Lecture notes of Prof. W. U. De

Leon
• Training Course in Diagnostic
Medical Parasitology –
Entomology module, 2000
• Study Guide in Medical
Parasitology, 1997
Thank you for listening!