SECTION 1 PART 2 Flashcards
1
Q
is essential for the isolation and proper diagnosis of infection.
A
Proper collection of fecal sample
2
Q
The following should be considered:
- Container: [?] (for physical examination of the specimen)
- Avoid contamination with [?]
- Label - [?]
A
sterile, disposable, wide mouth with tight-fitting lid, transparent
urine, water and soil
side of the container, not on the lid
3
Q
Submission of specimen:
- liquid
- formed
- watery
A
30 mins - liquid
1 hr - formed
15-30 mins - watery
4
Q
- Handle carefully because it is a potential source of infection unsuitable samples from patients receiving: UNSUITABLE SAMPLES FROM PATIENTS RECEIVING
A
a. Barium
b. Oil
c. Bismuth
d. Kaolin
e. Antibiotics
5
Q
TYPES of FECAL SPECIMEN
A
- Liquid
- Soft
- Formed
6
Q
– either diarrheic or saline-purged
A
- Liquid
7
Q
: process of administering saline that are ways to promote evaporation of watery stool
A
saline-purged
8
Q
a) Physical PRESERVATION
A
- Room temperature
- Refrigeration
9
Q
b) Chemical PRESERVATION
A
- Polyvinyl Alcohol (PVA)
- 5-10% formalin
- Merthiolate-Iodine-Formalin (MIF) or Thimerosal Iodine Formalin (TIF)
- Schaudinn’s Fixative
- Sodium Acetate - Acetic Acid Formalin (SAF)
10
Q
Refrigeration (?)
A
2 to 8 degrees; only formed to semi-formed
11
Q
used for the preservation of stained fecal+ smears
A
Polyvinyl Alcohol (PVA)
12
Q
aka Modified Schaudinn’s (with fixative)
A
Polyvinyl Alcohol (PVA)
13
Q
Polyvinyl Alcohol (PVA) components:
A
schaudinn’s fluid: 93.5 ml
glycerol: 1.5 ml
glacial acetic acid: 5 ml
polyvinyl alcohol (powder): 5 g
14
Q
*Quality control: discard if cloudy
A
Polyvinyl Alcohol (PVA)
15
Q
For concentration techniques or direct fecal smear
A
5-10% formalin
16
Q
For cysts, helminthes and larvae
A
5-10% formalin
17
Q
5-10% formalin
cysts:
helminthes and larvae:
stock solution:
A
cysts: 5%
helminthes and larvae: 10%
stock solution: 40%
18
Q
- may or may not have iodine
A
Direct fecal smear
19
Q
Not sufficient for preparing permanent stained fecal smears
A
5-10% formalin
20
Q
*Quality control: Heat at 60°C before use (temperature favors development of helminth)
A
5-10% formalin
21
Q
Merthiolate-Iodine-Formalin (MIF) or Thimerosal Iodine Formalin (TIF)
For bulk feces, the solution is mixed in the proportion of (?) and (?) for a gram of fecal material
A
9.4 ml MIF
0.6 ml Lugol’s
22
Q
For fresh materials and those recovered from intestinal mucosal linings
A
Schaudinn’s Fixative
23
Q
For permanent staining
A
Schaudinn’s Fixative
24
Q
without mercuric chloride; lipid fixative with a long shelf-life
A
Sodium Acetate - Acetic Acid Formalin (SAF)
25
image of the organism if not sharp after staining
Sodium Acetate - Acetic Acid Formalin (SAF)
26
IMPORTANCE OF FECALYSIS
1. To detect the presence of [?]
2. For the detection of evidence of malfunction of some parts of the [?]
3. For the detection of [?]
4. For the detection of [?]
5. Used as a clue in [?]
intestinal parasites
GIT, liver and pancreas
GIT bleeding
excessive fats in stool (steatorrhea)
medical and surgical diagnosis.
27
Liver damage: [?] gives the normal brown color.
Stercobilin
28
may indicate billiary obstruction
Acholic (gray/no color)
29
- upper GIT bleeding (red)
Hematochezia
30
- black
Melena
31
Form and Consistency Normal:
soft to formed
32
Form and Consistency Variations:
33
seen in diarrhea and administration of saline cathartic
a) Very soft and watery
34
px taking taxatives
a) Very soft and watery
35
constipation due to lack of mucus
b) Excessively hard and scybalous
36
: goat-droppings
scybalous
37
cholera
c) Rice water stool
38
- Does not flow on tilting due to trapped gas
Mushy
39
Sprue of excessive carbohydrate fermentation
Mushy
40
- liquid and consist mostly of water
Diarrheic
41
early typhoid
d) Pea soup stool
42
syphilis, spastic colitis and obstruction at the lower portion of the colon (rectum or anus)
e) Flattened or ribbon like
43
fibrocystic disease of the pancreas
f) Butter like
44
excessive carbohydrate fermentation
g) Gaseous and fermentative
45
Color Normal:
light brown to dark brown due to stercobilin
46
Color Variations:
47
Oxidized [?] gives off bilirubin
stercolobilinogen
48
due to administration of santonin (antihelminthic) and senna (antihelminthic or pang purge from plant)
a) Yellow
49
seen after ingestion of Barium meals
b) Light clay or putty color (acholic/colorless)
50
(used before x-ray)
Barium
51
bleeding in the lower GIT (hematochezia)
c) Reddish or bloody
52
undigested beets and tomatoes
c) Reddish or bloody
53
bleeding in the upper GIT
d) Dark red/chocolate brown
54
intense intake of coffee, chocolate, cherries, and black berry
d) Dark red/chocolate
55
associated with digestion of blood due to bleeding in the upper GIT
e) Black/tarry
56
increased intake of bismuth and charcoal
e) Black/tarry
57
amoebiasis
f) Greenish
58
mucoid
f) Greenish
59
fishy order
f) Greenish
60
vegetable ingestion (green stool w/o amoebiasis)
f) Greenish
61
calomel, mercury, chloride
f) Greenish
62
cocoa and chocolate produce dark a stool
g) Gray
63
infants (normal) and adults (lack of bilirubin derivatives)
h) Golden yellow/yellowishwhite
64
bizarre colors, whitish discoloration, blue and orange red
i) Miscellaneous
65
(drug-intake)
bizarre colors
66
blue –
methylene blue/dithiozaline
67
orange-red –
pyridium
68
Odor Normal:
foul to offensive due to indole, skatole and butyric acid
69
Abnormal odors:
70
found in ulcerative and malignant tumor of the lower bowel
a) Putrid odor
71
indicates gas formation, fermentation of carbohydrate, unabsorbed fatty acids
b) Sour/rancid odor
72
usually in alkaline stools, putrefaction of undigested protein.
c) Extremely foul odor
73
antibiotic intake kills both pathogenic and normal flora causing alkaline ph of stool
c) Extremely foul odor
74
– not routine in the lab; blood not visible to the naked eye and only detected by chemical means
Occult blood
75
Laboratory Examination for Occult Blood:
a) Benzidine test
b) Guaiac;s test
c) Hematest
76
Laboratory Examination for Occult Blood - most commonly used
Guaiac;s test
77
General Principle: The peroxidase-like activity of blood (due to peroxidase contained in the heme portion of hemoglobin) decomposes [?] to produce [?]. The liberation of [?] causes the oxidation of the colorless chromogen into a colored compound (blue)
H2O2
water and oxygen
oxygen
78
Patient Preparation: meat free diet for [?] prior to the test
3-5 days
79
Cause False (+) Results
1. Peroxidase activity substance or fecal material
2. Iron in the diet
3. Myoglobin in ingested meat
80
px should not eat:
melon, fresh uncooked broccoli, horse radish, cauliflower, and turnip
81
Cause False (-) Results
1. Large amount of Vitamin C
2. Breakdown of blood and its constituents
3. Hemorrhage in the Upper GIT 4, Technical errors (personnel, reagent problem or quality control)
82
strong reducing agent that blocks the activity of Guaiac, Hematest, and Benzidine
Vitamin C
83
The following structures may be seen microscopically:
84
– seen in watery or diarrheic sample
a) Trophozoites and cysts of amoeba
85
– seen in formed stool
b) Helminth eggs and larvae
86
– depends on diet
g) Plant cells, pollen grain and spores
87
– from eosinophil and basophil; evidence of parasitic infection
l) Charcot-layden crystals
88
TECHNIQUES
1. DIRECT FECAL SMEAR
2. KATO THICK SMEAR (KTS)
3. CONCENTRATION TECHNIQUES
4. STAINING METHODS
5. SPECIAL RECOVERY METHODS
6. CULTURE METHODS
7. ANIMAL INOCULATION TESTS
8. XENODIAGNOSIS
89
– simplest and most frequently used
DIRECT FECAL SMEAR
90
- for the observation of the motility of trophozoites
a) 0.85% sodium chloride/NSS
91
is used if there is no stain used
Normal saline
92
– for protozoan cysts and helminth ova
b) D’antonis solution/Lugol’s Iodine
93
• For large scale examination
DIRECT FECAL SMEAR
94
• Satisfactory for all kinds of helminth eggs
DIRECT FECAL SMEAR
95
•Unsuitable for diarrheic stools
DIRECT FECAL SMEAR
96
•Unsuitable for protozoan cysts and trophozoites
DIRECT FECAL SMEAR
97
- reagent used kills cysts and thropozoites that will make them look like contaminants
DIRECT FECAL SMEAR
98
DIRECT FECAL SMEAR Reagents:
a) 0.85% sodium chloride/NSS
b) D’antonis solution/Lugol’s Iodine
99
KATO THICK SMEAR (KTS) Reagents:
Distilled water 100 ml
Glycerin 100 ml
3% malachite green 1 ml
100
- green cellophane was formerly used; to prevent eye strain
3% malachite green
101
Use cellophane as cover slip
KATO THICK SMEAR (KTS)
102
KATO THICK SMEAR (KTS) Clearing time:
1 hour
103
- used in the detection of a small number of parasites which are not detected using DFS
CONCENTRATION TECHNIQUES
104
– uses either specific gravity or centrifugation
a) Sedimentation Method
105
a) Sedimentation Method Types
106
sediment - settles at the bottom; for all types of eggs especially operculated (with cap)
a) Sedimentation Method
107
– concentrates helminth eggs, larvae and protozoan cysts
• Formalin-Ether techniques
108
: fixative
formalin
109
: removes lipids to clear the sediments
ether
110
– used for formed stools; applicable for helminth eggs and larvae
• Acid-Ether Sedimentation
111
: clearing agent
Acid-ether
112
– time-consuming
• Simple sedimentation
113
- for concentration of microfilariae’ utilizes venous blood as specimen
• Knott Concentration Technique
114
To diagnose elephantiasis
• Knott Concentration Technique
115
- settles above; not suitable for operculated ova (D. Latum - tapeworm; Fasciola - liver flukes)
b) Flotation method
116
✓ Allows the separation of helminth eggs, protozoan cysts and larvae (1.05 – 1.15) using chemical solutions of higher specific gravity (1.12-1.21)
b) Flotation method
117
✓ Eggs and cysts float to the surface of the solution while fecal materials sink to the bottom
b) Flotation method
118
✓ Superior to sedimentation for concentrating cysts and eggs other than operculated, schistosomal and infertile Ascaris eggs
b) Flotation method
119
b) Flotation method Optimal time:
5 to 20 mins (after 30 minutes destroys/disintegrates the cysts)
120
- valuable method for concentrating cysts and eggs
• Zinc Sulfate centrifugal flotation technique
121
• Zinc Sulfate centrifugal flotation technique
- specific gravity:
[?] (hydrometer)
[?] formalinized feces
1.18
1:20
122
- Cryptosporidium (rounded oocysts)
• Sugar Flotation Technique/Sheather’s Sugar Flotation
123
• Sugar Flotation Technique/Sheather’s Sugar Flotation COMPOSITION:
Sucrose (500g), Tapwater (320 mL), Phenol (6.5g)
124
: to prevent mold formation
- Phenol
125
- uses sodium chloride solution
• Wilie’s Brine
126
• Wilie’s Brine COMPOSITION:
NaCl (40g) + Distilled water (100mL)
127
4. STAINING METHODS
128
• 1% Isotonic Eosin Solution & 2% Brilliant Cresyl Blue
- Trophozoites-
- Background-
- Trophozoites- blue green
- Background- pink
129
• D’Antonis
• 1% Isotonic Eosin Solution & 2% Brilliant Cresyl Blue
• Eosine in saline
• Buffered Methylene Blue (ex. Nairs Methylene Blue)
130
b) Permanent Staining
• Gomori’s Trichrome Stain
• Lawless’ Permanent Mount Stain
• Modified Acid-Fast stain
• Iron Hematoxylin Stain
• MIF Fixative stain ( Merthiolate iodine formaldehyde)*
• Chlorazol Black E*
• Modified Kohn’s*
131
- Wheatley’s modification
• Gomori’s Trichrome Stain
132
- For intestinal protozoa
• Gomori’s Trichrome Stain
133
- Rapid method of staining trophozoites and cysts
• Lawless’ Permanent Mount Stain:
134
• Lawless’ Permanent Mount Stain
- Protozoa –
blue to purplish color
135
- Cryptosporidium, Isospora and Cyclospora
• Modified Acid-Fast stain
136
- Two separate stains: Carbol fuchsin and methylene blue
• Modified Acid-Fast stain
137
- lacks specificity
• Modified Acid-Fast stain
138
- diagnosis of Trichomonas
• MIF Fixative stain ( Merthiolate iodine formaldehyde)*
139
- An acid dye, used as a fat and general tissue stain, and to stain protozoa in fecal smears or in tissues.
• Chlorazol Black E*
140
- modification of the chlorazol black E staining technique
• Modified Kohn’s*
141
5. SPECIAL RECOVERY METHODS
a) Cellulose Tape Preparation/Graham Scotch Tape Method
b) Egg Count Technique
c) Egg Hatching Technique
142
b) Egg Count Technique Methods
1. Direct Smear Egg Count (Method by Beaver)
2. Dilution Egg Count
3. Thick Smear Egg Count
4. Dilution-Filtration Egg Count
143
✓ Enterobius vermicularis and Taenia
Cellulose Tape Preparation/Graham Scotch Tape Method
144
✓ Collection: morning before the patient washes or defecates
Cellulose Tape Preparation/Graham Scotch Tape Method
145
✓ Commercial collection kits are available
Cellulose Tape Preparation/Graham Scotch Tape Method
146
Tongue depressor attached w/ glass slide and scotch tape
Cellulose Tape Preparation/Graham Scotch Tape Method
147
– provides an estimate of worm burden
Egg Count Technique
148
✓ Determines the degree of infection (light, moderate or heavy)
Egg Count Technique
149
✓ For recovery of hookworms, Ascaris and Trichuris
Egg Count Technique
150
1. Direct Smear Egg Count (Method by Beaver)
- 1.5 mg feces (?) = eggs per gram (epg)
- 2.0 mg feces (?) = epg
667
500
151
– uses 0.1 N NaOH - Stoll’s Egg Counting Technique
2. Dilution Egg Count
152
– KTS for schistosomes
3. Thick Smear Egg Count
153
– Schistosomes
4. Dilution-Filtration Egg Count
154
is a laboratory tool used to determine a given parasite's resistance to extant drug therapy.
Egg Hatching Technique
155
CULTURE METHODS PURPOSES:
✓ For accurate detection of the organisms as a [?] to other methods
✓ For obtaining a rich yield of organisms to be used as [?] in immunologic diagnosis [?]
✓ For in-vitro screening of [?]
✓ For investigating the [?] of organisms
✓ As a source for inoculating [?]
supplement
antigen
drugs
physiology
susceptible experimental animals
156
CULTURE METHODS for PROTOZOA
a) Balamuth’s Monophasic Medium
b) Boeck & Drbohlav’s Diphasic as Modified by Dobell and Laidlaw
c) Cleveland & Collier’s
d) Trussel and Johnson’s medium (& Fineburg and Whittington)
e) Axenic Culture Method
157
- Entamoeba histolytica and Balantidium coli
Balamuth’s Monophasic Medium
158
- E. histolytica
Boeck & Drbohlav’s Diphasic as Modified by Dobell and Laidlaw
159
- Concentrate from saline centrifugal sedimentation of feces rather than the feces is recommended.
Boeck & Drbohlav’s Diphasic as Modified by Dobell and Laidlaw
160
- Use of whole hen’s eggs
Boeck & Drbohlav’s Diphasic as Modified by Dobell and Laidlaw
161
- highly specific for E. histolytica
Cleveland & Collier’s
162
Isolation of Trichomonas vaginalis
Trussel and Johnson’s medium (& Fineburg and Whittington)
163
- histolytica
Axenic Culture Method
164
CULTURE METHOD FOR HOOKWORMS AND STRONGYLOIDES
A. Coproculture
B. Harada-Mori or Test tube culture method
165
CULTURE METHOD for LEISHMANIA & TRYPANOSOMES
a) Novy, MacNeal and Nicole (NNN) Diphasic
b) Cellular Media
166
- (+) stool samples are mixed with moisture soil/granulated charcoal for the development of larvae
Coproculture
167
- BAERMANN METHOD or funnel method
A. Coproculture
168
- (+) positive stool sample is applied to the filter paper, and immersed with boiled distilled water
B. Harada-Mori or Test tube culture method
169
Strongyloides (strong) follows an upward movement unlike hookworm that goes downward
B. Harada-Mori or Test tube culture method
170
- Best medium
Novy, MacNeal and Nicole (NNN) Diphasic
171
- For Trypanosoma cruzi and Toxoplasma
Cellular Media
172
• RBCs that have been coated with antigen are allowed to react with the test serum
Indirect Hemagglutinat ion (IHA)
173
• The red cells agglutinate in the presence of specific antibady
Indirect Hemagglutinat ion (IHA)
174
• Bentonite, latex particles or the organism is exposed to the test serum
Flocculation or Agglutination
175
• Agglutination indicates the presence of a specific antibody
Flocculation or Agglutination
176
• Microscopically visible parasites fixed on a slide are allowed to react with test serum
Indirect Fluorescent Antibody (IFA)
177
• The slide is washed and treated with AHG with fluorescein (Sandwich Method)
Indirect Fluorescent Antibody (IFA)
178
– very sensitive in several cases; requires concentrated
antigen
Gel Diffusion(Ouchterlony) and Countercurent Electrophoresis (CEP)
179
• Test serum is placed in a well adjacent to the antigen
Gel Diffusion(Ouchterlony) and Countercurent Electrophoresis (CEP)
180
• Both reactant are allowed to diffuse toward each other through the agar
Gel Diffusion(Ouchterlony) and Countercurent Electrophoresis (CEP)
181
• Formation of precipitates/band between the
Gel Diffusion(Ouchterlony) and Countercurent Electrophoresis (CEP)
182
- color reaction is measured spectrophotometrically
Enzyme linked immunosorbe nt assay (ELISA)
183
• Test serum is allowed to react with an antigen (coat the surface of plastic well).
Enzyme linked immunosorbe nt assay (ELISA)
184
• Anti-human IgG antibody conjugated with enzyme is added.
Enzyme linked immunosorbe nt assay (ELISA)
185
• If a specific antibody was present in the test serum, the anti-human IgG antibody labeled with an enzyme attaches to the antigenantibody complex.
Enzyme linked immunosorbe nt assay (ELISA)
186
The addition of a suitable substrate generates a visible color reaction
Enzyme linked immunosorbe nt assay (ELISA)
187
Enzyme linked immunosorbe nt assay (ELISA) spp
T. gondii, T. vaginalis, leishmaniasis, malaria, schistosomiasis etc.
188
✓ Animal Used: hamster
Leishmania
189
✓ Specimen: aspirates from biopsy materials from cutaneous ulcers, lymph nodes, spleen, liver
Leishmania
190
Leishmania Specimen Collection
intravenous, intramuscular, subcutaneous, intradermal, intraperitoneal, intracerebral, intranasal
191
✓ Animal Used: Guinea pigs or white rats (Trypanosoma gambiense and T. rhodesiense)
Trypanosoma
192
✓ White mice (?)
Trypanosoma cruzi
193
✓ An experimental bug (vector) is allowed to take a blood meal from the patient
XENODIAGNOSIS
194
✓ The intestinal contents of the bug is assessed for the presence of diagnostic stages of the parasite
XENODIAGNOSIS
195
XENODIAGNOSIS Ex. T. cruzi from an
assassin bug or kissing bug
196
: The trichrome technique of Wheatley for fecal specimens is a modification of Gomori's original staining procedure for tissue.
Gomori’s Trichrome Stain
197
It is a rapid, simple procedure which produces uniformly well stained smears of the intestinal protozoa, human cells, yeast cells, and artifact material in about 45 min or less.
Gomori’s Trichrome Stain
198
: A rapid permanent-mount stain technic for the diagnosis of the intestinal protozoa
Lawless’ Permanent Mount Stain
199
Use of the modified Ziehl-Neelsen stain for faecal smears has already been established for coccidian protozoa, in particular, oocysts of Cryptosporidium species, but it is also useful to confirm the presence of oocysts of Isospora belli and Cyclospora cayetanensis
• Modified Acid-Fast stain
200
- used for most of the original morphological descriptions of intestinal protozoa found in humans.
• Iron Hematoxylin Stain
201
- On oil immersion power (1,000 x ), one can examine the diagnostic features used to identify the protozoan parasite.
• Iron Hematoxylin Stain
202
can be used with either fresh, SAFpreserved, or PVA-preserved specimens.
• Iron Hematoxylin Stain
203
✓ Fresh eggs are incubated from the parasite of interest and serial dilutions of the drug of interest are applied.
Egg Hatching Technique
204
The percentage of eggs that hatch or die is determined at each concentration and a drug response curve may be plotted.
Egg Hatching Technique
205
The data can then be transformed and analysed to give further statistics such as an ED50.
Egg Hatching Technique
206
✓ This technique is labour intensive, expensive and can take some time, however an egg hatch assay will give and accurate and reliable
Egg Hatching Technique
207
- All liquid medium commonly used for the isolation and maintenance of Entamoeba histolytica and some other intestinal protozoa.
Balamuth’s Monophasic Medium
208
- It is satisfactory for Balantidium coli, although the quality and amount of starch may need to be adjusted
Balamuth’s Monophasic Medium
209
- Use of egg yolk
Balamuth’s Monophasic Medium
210
- Egg slant
Boeck & Drbohlav’s Diphasic as Modified by Dobell and Laidlaw
211
- highly specific for E. histolytica , which grows luxuriantly on this medium.
Cleveland & Collier’s
212
Other intestinal amoebae do not grow readily on this medium.
Cleveland & Collier’s
213
The technique of overlaying the medium with fresh sterile horse serum-saline mixture, as reported by
Cleveland & Collier’s
214
was reported to be the best method of isolation of E. histolytica .
Cleveland & Collier’s
215
- Proteose peptone and infusion from liver provide amino acids and other nitrogenous substances that support growth of E. histolytica .
Cleveland & Collier’s
216
- Sodium chloride maintains the osmotic balance of the medium and sodium a-glycerophosphate acts as a phosphorous source.
Cleveland & Collier’s
217
has been an essential component of numerous immunological and biochemical studies
Axenic Culture Method
218
- Monophasic clear liquid for rapid growth of dense populations of amebae
Axenic Culture Method
219
- Use of sterile defibrinated rabbit blood
Novy, MacNeal and Nicole (NNN) Diphasic
220
amoebiasis, Chaga’s disease, malaria, toxoplasmosis, cystisercosis, hydatid cyst, filariasis, schistosomiasis
Indirect Hemagglutinat ion (IHA)
221
Leishmaniasis, Chaga’s disease
Flocculation or Agglutination
222
Detect amebiasis and fascioliasis infections
Gel Diffusion (Ouchterlony) and Countercurent Electrophoresi s (CEP)
223
✓ Modification of the standard ELISA for increased sensitivity and specificity in detecting antibodies of the IgM class to Toxoplasma
Antibody Capture or Double Sandwich ELISA
224
✓ Plastic well are coated with anti-human IgM antibodies
Antibody Capture or Double Sandwich ELISA
225
✓ After washing, the test serum is added followed by further washing and addition of the test antigen
Antibody Capture or Double Sandwich ELISA
226
✓ The final reagent is an antigen-specific antibody to which the color reagent has been conjugated
Antibody Capture or Double Sandwich ELISA
227
✓ The plates are read spectrophotometrically
Antibody Capture or Double Sandwich ELISA
228
✓ This method can be used for assay of parasitespecific IgE antibodies by starting with anti human IgE
Antibody Capture or Double Sandwich ELISA
229
is based on the fact that live Toxoplasma tachyzoites can actively take up methylene blue dye from the culture medium, whereas parasites that are killed because of complement-mediated lysis do not take up the dye and remain colorless.
SabinFeldman Dye Test
230
- Specifically designed for the diagnosis of Toxoplasmosis
SabinFeldman Dye Test
231
is a serological test used for diagnosis of schistosomiasis japonica.
Circumoval Precipitin Test (COPT)
232
formation of the circumoval precipitin by serum from infected humans.
Circumoval Precipitin Test (COPT) Soluble egg antigens of Schistosoma japonicum block the
233
was used to monitor purification of the soluble egg antigens of S. japonicum.
circumoval precipitin inhibition
234
is a very sensitive in vitro assay technique used to measure concentrations of antigens by use of antibodies.
Radioimmuno assay (RIA)
235
As such, it can be seen as the inverse of a radiobinding assay, which quantifies an antibody by use of corresponding antigens
Radioimmuno assay (RIA)
236
is a radioimmunoassay test to detect specific IgE antibodies to suspected or known allergens for the purpose of guiding a diagnosis about allergy
Radioallergosorbent Test (RAST)
237
- In this test, the parasite antigen is first bound to an inert sorbent or matrix to which the test serum is allowed to react.
Radioallergosorbent Test (RAST)
238
After washing, radio-labeled antibody (IgE or IgG) is added to the sorbent.
Radioallergosorbent Test (RAST)
239
After another washing to remove excess labeled antibody, the remaining radioactivity is a measure of antigen-specific IgG or IgE in the test serum
Radioallergosorbent Test (RAST)
240
CSF and serum may be used.
Immunoblotting/Western Blot
241
By using electrophoresis technique, the DNA containing portion of the patient sample is transferred to a polyacrylamide gel.
Immunoblotting/Western Blot
242
This process results in the separation of protein antigens in the gel.
Immunoblotting/Western Blot
243
A blotting technique is employed to transport the proteins onto a paper.
Immunoblotting/Western Blot
244
Giardiasis
Immunoblotting/Western Blot
245
Three ingredients is necessary to for performing the first phase: patient serum, a commercial antigen source, known amount of complement.
Complement Fixation (CF)
246
The complement is “fixed” by the antibodies present in the serum.
Complement Fixation (CF)
247
In the 2nd phase, sensitized sheep’s rbc is added to the mixture.
Complement Fixation (CF)
248
The absence of free complement prevents lysis of these cells ( positive). If the patient sera does not contain Ab, complement is free to lyse ( negative)
Complement Fixation (CF)
249
leishmaniasis, Chaga’s disease, pneumocystosis, paragonimiasis
Complement Fixation (CF)
250
Over the years, parasites which were once considered commensal have evolved to become
human pathogens
251
During this time, tremendous knowledge was obtained of the epidemiology, parasite-host relationships, life cycles, disease processes and symptoms, treatment, and prevention and control of
parasites
252
In addition, parasites are classified based on their individual
characteristics
253
Traditional as well as new methodologies for parasite identification allow for
accurate laboratory diagnosis
254
is an interesting and exciting field of the clinical laboratory sciences.
Parasitology
255
The continued development of high-tech, highly sensitive[?] provides the key to the future of parasitology.
parasite test methodologies
256
Because it is highly unlikely that parasites will totally be eradicated in the near future, competent practitioners educated in the field of parasitology are essential to ensure proper
parasite identification.
257
preservation of trophozoites and cysts
Polyvinyl Alcohol (PVA)
