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42
STOOL EXAMINATION
•Culture
•Consistency •Cellophane tape
•Colour Permanent Temprory •Baeremann tech.
•Composition •Ova quantitaion (Stoll & Kato)
Floatation
Sedimentation
43
WET MOUNT PREPARATION
It may be more cost effective to delete the direct smear and begin the
stool examination with the concentration procedure.
46
STOOL EXAMINATION
Temporary
Saline smear Iodine smear
saline Iodine 1%
•It can only be performed on fresh •Contain an excess of fecal debris that
material or on specimen preserved in could mask the presence of parasitic
preservatives containing formalin cysts.
STOOL EXAMINATION
Scanty infection
Concentration techniques
Sedimentation Floatation
• Non Operculated eggs
• Heavy eggs (Ascaris
egg) Trematodes ( S. m.)
Saline Emulsify
Conical flask
10 g stool
Sediment
52
STOOL EXAMINATION
Formol Ether Sed. Conc.
Ether
Ether debris
1 g stool
Sediment
Thorough mixing Conical flask centrif. tube
Tin container
• Cestode eggs (non op) Seive
•Egg of S.m.
•Nematode eggs?????
•Eggs of small tapeworms • Crypto, Iso. oocysts
•Hookworms???????
•Cysts
•Trichostong؟؟؟؟؟؟؟؟؟؟؟
20 min Centrif. 2 min
54
STOOL EXAMINATION
Kato technique
Mesh screen
Hole
Template
Remove the
template
Cellophane soaked by
glycerin (clears faeces(
24 hr stool
60 CC
4 g Stool
56 CC
Shake well 0.15 CC
NaOH
Egg count/ slide
Eggs/1g= Eggs/slideX
Erlynmeyer flask Egg/day=Eggs/1g
56 X stool
STOOL EXAMINATION
Baermann’s technique
Stool/soil
seive
25-50 CC
Warm water
Glass funnel 30 min centrifuge
clamp
Filter paper
Slide
Sealed petri dish
Water
Scanty infection
Larvae of:
• St. stercoralis (A,L)
• Hookworms
59 • Trichostrong
INDIRECT IMMUNOLOGICAL
METHODS
• Scanty infection.
• Tissue parasite no portal of exit (Hydatid
dis.)
• Migratory stage (Fasciola)
• Chronic infection fibrosis (Bilharziasis)
60
STOOL EXAMINATION
Permanent Stained smears
61
INDIRECT IMMUNOLOGICAL METHODS
ore specific
ore accurate.
ctive infection
Ab remain in serum for
arly
months even after cure
uantitative
62
Antigen detection
IHAT LAT
Ag
Ag +
+
Latex particlePatient’s serum
Patient’s serum (?? AB)
Sensitized
heep’s RBC (?? AB)
(O–ve)
65
Agglutination Agglutination
Immunofluorescent-antibody test (IFAT)
using monoclonal probes specific for Enterocytozoon bieneusi or
Encephalitozoon intestinalis.
• The sensitivity and specificity of IFAT were 100% Moreover,
• Secies identification by IFAT was more rapid and less expensive
than that by PCR.
• IFAT is a suitable test for detection of microsporidia in
developing countries.
INDIRECT IMMUNOLOGICAL METHODS
INDIRECT FLUORESCENT ANTIBODY TEST
fluorescein
Anti human AB
Patient’s serum
(?? AB)
parasite
67
INDIRECT IMMUNOLOGICAL METHODS
ELISA
OPD
Peroxidase E OPD
Anti human AB
atient’s serum
(?? AB) AB
Ag
Flat bottom plastic micrititre plate
68
INDIRECT IMMUNOLOGICAL METHODS
CFT
Sheep’s
Anti sheepRBC
AB
AB
comple
ment
Patient’s serum
(?? AB)
Ag
Tube /
microplate 69
INDIRECT IMMUNOLOGICAL METHODS
Double Electro Immuno Diffusion
Line of
ppt
Electric current
Ag A
b
Buffere
d gel
70
INDIRECT IMMUNOLOGICAL METHODS
Immunodiagnostic Strip Test (Dip Stick Test) Ag
Pt bld (?Ag)
Coloured dye
Monoclonal Ab
trocellulose strip Malaria, Filaria, African tryp.
PCR
Replication
73
Detection
T cruzi, T gondii
10 X Objective
74
40 X Objective
75
2 VIAL COLLECTION KITS
5% OR 10%
FORMALIN PVA
PERMENANT
STAINED SMEAR
ONCENTRATION
Trichrome or
Iron haematoxylin
GENERAL PRECISE
MORPHOLOGY MORPHOLOGY
Non-parasitic structures found in faeces:
Care
must be taken not to report as parasites those
structures that can be normally found in faeces such as:
muscle fibres, vegetable fibres, starch cells (stain blue-
black with iodine), pollen grains, fatty acid crystals,
soaps, spores, yeasts, and hairs .
Large numbers of fat globules may be seen in faeces
when there is malabsorption.
Charcot Leyden crystals (breakdown products of
eosinophils) can sometimes be seen in faeces (also in
sputum) in parasitic infections. They appear as slender
crystals with pointed ends, about 30–40m in length
77
Structures found in faeces that required
differentiation from parasites.
Image illustrating Yeast Cells in slide preparation79 Image illustrating Vegetable cell in slide
Note similarity to parasitic oocysts. preparation.
Image illustrating Vegetable Spiral in slide
preparation.
Image illustrating pollen in slide preparation Image illustrating geranium pollen cells in
81 slide preparation.
using a color filter
Image illustrating peach hair Image illustrating vegetable
in slide preparation. Note the hairs in slide preparation.
similarity to Strongyloides
stercoralis. 82