DIAGNOSTIC PARASITOLOGY
Francis Anthony V. Soriano, RMT
PAR 313 LEC
College of Medical Technology
Mt. Carmel College-Bocaue, Bulacan
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PARASITOLOGY Laboratory Diagnosis
-accurate diagnosis of parasitic infections can help
decrease the prevalence and incidence of a parasitic
infection.
1. Confirm clinical impression
2. Rule out diagnosis
3. Aid a clinician in the choice of proper medication
4. Help in monitoring the effect of treatment
regimen
Diagnostic parasitology is done by:
A. Demonstration of parasites (e.g., eggs, larvae,
adults, cysts, oocysts, trophozoites)
B. Detection of host immune response to the
parasites (e.g., Abs and Ags) 2
PARASITOLOGY
OUTLINE OF TECHINIQUES (stool)
1. Direct Fecal Smear
2. Kato-thick Smear
3. Concentration
a. Sedimentation Techniques
b. Floatation Techniques
4. Stool Culture
5. Egg counting procedures
6. Perianal Swab
7. Staining stool specimen
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PARASITOLOGY 1. DIRECT FECAL SMEAR (DFS)
• Routine method of stool examination
• Primarily useful in detection of motile protozoan
trophozoites
• Trophozoites are pale and transparent
• Nair’s buffered methylene blue (BMB) – demonstrate
nuclear morphology of trophozoites
• Lugol’s iodine – temporary stains nuclei of protozoan cysts
• Employs use of approx. 2 mg of stool
* Light infections may not be detected
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PARASITOLOGY 2. KATO THICK SMEAR
• Employs use of 50-60 mg of stool (size of two mongo beans)
• Uses cellophane paper soaked in a mixture of glycerine and
malachite green solution
• Simple and economical
• Very good in detecting eggs with thick shells (e.g., Ascaris
and Trichuris) but not thin shells
* Usefulness is limited in diarrheic and watery stools
* Not able to detect protozoan cyst and trophozoite
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PARASITOLOGY 3. CONCENTRATION TECHNIQUES
• Useful in cases of light infections
A. Sedimentation Procedures
1. Acid Ether Concentration Technique (AECT)
- 40% HCl→ dissolves albuminous material
- ether → dissolves neutral fats
- recommended for recovery Trichuris, Capillaria and
trematode eggs specially Schistosoma.
- Drawbacks: loss of parasite to the plug of debris and
possible destruction of protozoan cyst
2. Formalin Ether Concentration Technique (FECT)
- Formalin as preservative
- Ethyl acetate as alternative to ether
- more efficient in recovering cestode eggs and Giardia
cysts
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PARASITOLOGY
PARASITOLOGY 3. CONCENTRATION TECHNIQUES
• Useful in cases of light infections
B. Floatation Procedures
1. Zinc Sulfate (ZnSO4) Floatation
2. Brine Floatation
3. Sheather’s Sugar Floatation
-employs use of boiled sugar solution preserved with
phenol
- best for recovery of coccidian oocysts
(e.g., Cryptosporidium, Cyclosopa and Isospora)
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PARASITOLOGY 4. STOOL CULTURE METHODS
• Useful for hookworm identification
• Stools positive for hookworm ova and/or Strongyloides
rhabditiform larva can be cultured until the filariform larvae
develop
A. Coproculture
- positive stools mixed with moistened soil and
granulated charcoal
B. Harada-Mori or the Test tube Culture Method
- employs use of test tubes and filter paper strips
- also used for cultivation of intestinal protozoan
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PARASITOLOGY 5. EGG COUNTING PROCEDURES
• Used to correlate the severity of clinical disease with the
intensity of infection or worm burden
A. Kato-Katz Method or Cellophane Covered Thick Smear
B. Stoll Egg Count
WHO classification of intensity of infections with STH and
Schistosomiasis
Organism Light Intensity Moderate Intensity Heavy Intensity
A. lumbricoides 1-4,99 epg 5,000-49,999 epg ≥ 50,000 epg
T. trichiura 1-999 epg 1,000-9,999 epg ≥ 10,000 epg
Hookworm 1-1,999 epg 2,000-3,999 epg ≥ 4,000 epg
S. japonicum and 1-99 epg 100-399 epg ≥ 400 epg
S. mansoni
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PARASITOLOGY 5. EGG COUNTING PROCEDURES
A. KATO-KATZ METHOD or CELLOPHANE COVERED THICK
SMEAR
• Uses a measured amount of stool which has been sieved
through a wire mesh and pressed under cellophane paper
soaked in malachite green solution
• Uniform amount of stool is examined using a template with a
uniform sized hole in the middle
• Consistency is the main determinant of the sensitivity of this
technique
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PARASITOLOGY 5. EGG COUNTING PROCEDURES
B. STOLL EGG COUNT
• 0.1 N NaOH as stool diluent, it saponifies fat and free eggs
from fecal debris
• Uses Stoll pipettes calibrated at 0.075 mL and 0.15 mL
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PARASITOLOGY
PARASITOLOGY 6. PERIANAL SWAB
• Used to recover eggs of E. vermicularis and Taenia spp.
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PARASITOLOGY 7. STAINING OF STOOL SAMPLES
• Performed specifically for the examination of the nuclear
characteristics of amoeba.
• Also useful for ID of other intestinal protozoans such as
Balantidium coli and Giardia spp.
a. Iron- hematoxylin
b. Trichome
c. Chlorazol Black E
• Kinyoun staining for coccidians
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