Here is the slide on Laboratory diagnosis of Parasites

1. L A B O R A T O R Y
D I A G N O S I S
O F P A R A S I T I C
I N F E C T I O N
Nawang Sherpa
Kiran Sanjyal
Nikki Poudel
Rashmi Tamang
S T . X A V I E R ’ S C O L L E G E , N E P A L
B S C M I C R O B I O L O G Y F I R S T Y E A R

2. • When any Parasites live in any of its host, they noticeably may
not cause infection. Most of them are known to cause dreadful
disease but few of them live unnoticed.
• As per “Old Friend Hypothesis”, Parasites may be our old
friends who are benefitted by living in our body. It is always
important to know what kinds of Parasites live in our body.
• Whether, they cause serious disease or just a mere presence
in our body.
Here, We present to you our Presentation of How can we
diagnose for the Parasites living in our body

3. D I R E C T S P E C I M E N S U S E D
F O R D I A G N O S I S O F
PA R A S I T I C I N F E C T I O N S
• 1)Stool
• 2)Urine
• 3)Sputum
• 4)Biopsy material from spleen, liver and lymph node.
• 4)Bone Marrow
• 5)Blood

4. U S E O F S TA I N S I N PA R A S I T I C
I N F E C T I O N
• Stains can help diagnose all types of parasitic infections.
We're going to look at parasitic stains for protozoa, helminth.
Protozoal stain
-Giemsa stain for blood parasite:
stains color genetic material and cytoplasmic components of cells a blue to purple color with eosin dye
Helminth stain
-Trichrome stain, Iron hematoxylin stain, Modified acid-fast stain
for detection of larvae and eggs
Uses specimen(faeces) fixed with formalin

5. D I A G N O S I S O F P A R A S I T I C I N F E C T I O N S
T H R O U G H F A E C E S
• Macroscopic Examination
It includes Presence of blood, Consistency, Colour, Odour of Faeces.
Faeces with Toxocara cati
Ropeworm Infection

6. • Microscopic Examination
-easiest and simplest technique
-Wet mount of faecel material is
prepared by using Iodine or Saline.
In Saline Wet Mount Examination
-Preliminary examination
for detection of Trophozoites
or cysts and Helminth
larvae or eggs
In Iodine Wet Mouth Examination
-staining nuclei and
cytoplasm by using Iodine Enterobias vermicularis

11. All eggs can be observed by this method except unfertilized eggs of
few which floats

23. Fasciola hepaticaEntamoeba histolytica Giardia lambila

24. D I A G N O S I S O F P A R A S I T I C I N F E C T I O N S
T H R O U G H U R I N E
• The specimen is collected in a sedimentation of glass and the
eggs are allowed to settle down.
• A drop of sediment is placed on a glass slide along with the
eggs and examined for the presence of Trichomonas
vaginalis

26. D I A G N O S I S O F P A R A S I T I C I N F E C T I O N S
T H R O U G H S P U T U M
• Microscopic examination of sputum is used in identifying Paragonimus
westermani eggs, Strongyloides stercoralis larvae, Ascaris
lumbricoides larvae, hookworm larvae, and rarely Entamoeba histolytica.
The sputum is spread in a petridish and cultured. The
region suspected for eggs/cysts or Charcot Leyden
crystals are examined for microscopic examination
Paragonimus westermani

27. D I A G N O S I S O F P A R A S I T I C I N F E C T I O N S
T H R O U G H B L O O D
• Preparation of Thin Blood smear
-Prick a finger or ear lobe with surgical cutting needle under aseptic technique.
-Take blood smear not larger than pin’s head under a clean glass slide.
-Make a dry smear with spreader.
-Pour Giemsa stain and leave it for 2 minutes.
-Dilute it with twice the volume of Distilled Water and leave for 10-15 minutes.
-After it is dried, observe it in microscope for possible protozoan parasite

28. Leshmania donovani Plasmodium falciparum

29. • Preparation of Thick blood smear
-A big drop of blood is taken in a glass slide and spear with a corder of glass slide.
-The blood should be spread properly.
-The slide is allowed to dry by covering with petridish and keeping in incubator for short time.
-Afterawhile, The film is flooded with misture of glacial acetate and tartarate.
Then, fixed with Methyl alcohol for 3-5 minutes and washed in distilled water.
The Giemsa stain is applied.

30. Enzyme-linked immunoelectrotransfer blot Taeniasis

32. T Y P E S O F A — A B R E A C T I O N S
( I N V I T R O )
• Agglutination
• Precipitation
• Complement Fixation Test (CFT)
• Neutralization
• Immunofluorescence Test (IF)
• Enzyme Linked Immunosorbent Assay (ELISA)
• Radio Immunoassay (RIA)
• Immunochromatography(ICT)
• Fluorescent Antibody Test (FAT)
• Bentonite Floculation Test
• Enzyme-linked immunoelectrotransfer blot (EITB)

33. A G G L U T I N A T I O N
• -It is the viable clumping of particulate(insoluble) Ag with its specific Ab forming visible
lattice. The Ab is the divalent agglutinating one either IgG or IgM type.
• The reaction can be either on side or tube agglutination
• It can direct or via the use of carrier (Indirect)example Latex, R.B.C (haemmaglutination)
• Or Staph protein A (Coagulation), Indirect more easily visible.
• The reaction can be qualitatively or quantitatively expressed.
Widal test for diagnosis of Giardia
Lates Agglutination test for diagnosis of Toxoplasma gondii

37. Presence of Malarial parasite infer a strong positive Complement Positive test i.e., No haemolysis

39. Mostly used in diagnosis for Protozoan Parasite like Plasmodium,
Leishmania, etc which produces antigens acceptable for Neutralization test to
be positive

41. ELISA is the most widely used serodiagnostic method for detection of parasite. It can
be used to detect most of the protozoan and metazoan parasite by the use of enzymes
to detect antibodies in the blood

48. Giardia as viewed from IFA

58. E N Z Y M E - L I N K E D
I M M U N O E L E C T R O T R A N S F E R B L O T
( E I T B )
• Enzyme-linked immunoelectrotransfer blot (EITB) or immunoblot
- a test that combines the high sensitivity of the immunoenzymatic tests
with the high resolution of sodium dodecyl sulfate-polyacrylamide gel
electrophoresis (SDS-PAGE)
- this method has been successfully adapted for the confirmatory
serodiagnosis of various parasitic diseases in humans, such as
schistosomiasis, hydatidosis, cysticercosis, taeniasis, fascioliasis and
strongyloidiasis

59. M O L E C U L A R M E T H O D S F O R D E T E C T I O N
O F PA R A S I T E S T R U C T U R E S
• Polymerase Chain Reaction (PCR)
The effectiveness of PCR in detecting parasite species such as Leishmania and Plasmodium
has already been studied
• Real-Time Polymerase Chain Reaction (RT-PCR)
• application of SYBR Green I RT-PCR to protozoans, including species of Cryptosporidium,
Leishmania and Trypanosoma
• he RT-PCR technique was able to detect Giardia in sewage and also proved to be a sensitive
method for identifying this protozoan in samples of human feces

60. • Loop-Mediated Isothermal Amplification (LAMP)
the LAMP technique to detect several parasitic diseases, including the human
parasites Cryptosporidium, Entamoeba
histolytica, Plasmodium, Trypanosoma, Taenia, Schistosoma, Fasciola hepatica and Fasciola
gigantica, and Toxoplasma gondii, and animal parasites such as Theileria and Babesia
• Random Amplified Polymorphic DNA (RAPD)
Studies using this technique show that it is able to differentiate species of Leishmania, in addition to
being used to study polymorphisms of parasites of medical importance such
as Plasmodium and Trypanosoma
• Microsatellites
microsatellite markers were developed only for some parasitic nematodes such as species
of Trichostrongyloid nematode