Lab Diagnosis of Parasitic Infection - yimg.com

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21 Φεβ 2014 (πριν από 3 χρόνια και 4 μήνες)

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LABORATORY
DIAGNOSIS

OF

PARASITIC

INFECTIONS

Case diagnosis


History (Age, occupation, residency, previous
infection)


Complaint


Clinical examination


Invesigations


-

Laboratory investigations


-

Radiology


-

Surgical intervention (Exploratory)


Provisional diagnosis

Confirm the diagnosis

DIAGNOSIS

DIRECT

INDIRECT

MOLECULAR

Urine

Stool

Sputum

Biopsy

Blood

Aspirates

PCR

DNA probes

IHAT

LAT

IFAT

ELISA

CFT

DEIDT

URINE EXAMINATION

MACROSCOPIC

MICROSCOPIC

colour

white

smoky

Sedimentation

concentration

Membrane filtration

Chyluria

Blood

Filaria

S. haematobium

Acetic acid



RBC haemolysis


Clear ova

Ether


Dissolve fat


M.f

URINE EXAMINATION

SEDIMENTATION



CONCENTRATION

15
-
20
min

Centrifuge (
2
min)

Clean conical
glass receptacle

URINE EXAMINATION

Membrane filtration technique

air

10
ml urine

Nucleopore filter

Eggs of Schistosoma

+ Saline

URINE EXAMINATION

HELMINTHES

PROTOZOA

ARTHROPODES



S. haem.egg



E. vermic. egg



S. mansoni egg



Mf (Ov, Wb)



H sand



T. Vag troph



Pthirus pubis



L. higher deptera

URINE EXAMINATION

Egg viability

Live eggs

Dead eggs


Well defined miracidium


Flickering F cells


Hatching moving miracidium



Dark colour


Granulated

STOOL EXAMINATION

MACROSCOPIC

MICROSCOPIC

OTHERS


Consistency


Colour


Composition


Culture


Cellophane tape


Baeremann tech.


Ova quantitaion (Stoll & Kato)


Temprory

Permanent

Diect saline smear

Iodine smear

Concentration techniques

Sedimentation

Floatation

Saline

Formol ether

Sat

saline

Zinc sulphate

Sheather

s sugar

MACROSCOPIC EXAMINATION

COLOUR

CONSISTENCY

COMPOSITION

Adult PARASITES

Pale=Steatorrhea


(G.l)

-
Liquid (Troph)

-
Formed (Cyst)

-
Semi formed (Cyst)

?? Blood ?? Mucus

(dysentry)

*Ascaris worm

*E. vermicularis

*T. saginata

STOOL EXAMINATION

STOOL EXAMINATION

MACROSCOPIC

MICROSCOPIC

OTHERS


Consistency


Colour


Composition


Culture


Cellophane tape


Baeremann tech.


Ova quantitaion


(Stoll & Kato)


Temprory

Permanent

Diect saline smear

Iodine smear

Concentration techniques

Sedimentation

Floatation

Saline

Formol ether

Sat

saline

Zinc sulphate

Sheather

s sugar

STOOL EXAMINATION

Temporary

Saline smear

Iodine smear


saline

Iodine
1
%

Huge number of:


Eggs



Protozoal troph. Motility

(Amoeb, flagellates)

Huge number of:


Cyst morphological details

STOOL EXAMINATION

Scanty infection

Concentration techniques

Sedimentation

Floatation



Heavy eggs (Ascaris egg)



Operculated eggs (Trematodes)



Larvae (Strong sterc.)



Cysts





Non Operculated eggs


Trematodes ( S. m.)


Cestode


Nematode(
Hookworms,Trichostong)



Cysts

STOOL EXAMINATION

MACROSCOPIC

MICROSCOPIC

OTHERS


Consistency


Colour


Composition


Culture


Cellophane tape


Baeremann tech.


Ova quantitaion


(Stoll & Kato)


Temprory

Permanent

Diect saline smear

Iodine smear

Concentration techniques

Sedimentation

Floatation

Saline

Formol ether

Sat

saline

Zinc sulphate

Sheather

s sugar

STOOL EXAMINATION

Saline sedimentation


10
g stool

Saline

Mesh wire gauze

Conical flask

Sediment

Emulsify

STOOL EXAMINATION


Formol Ether Sed. Conc.


10
% Formalin

1
g stool

Sediment

formalin

debris

Ether

Thorough mixing

Ether



Ether adsorbs fecal debris & floats.



Formalin fixes & preserves the specimen.

Conical flask centrif. tube

STOOL EXAMINATION

MACROSCOPIC

MICROSCOPIC

OTHERS


Consistency


Colour


Composition


Culture


Cellophane tape


Baeremann tech.


Ova quantitaion


(Stoll & Kato)


Temprory

Permanent

Diect saline smear

Iodine smear

Concentration techniques

Sedimentation

Floatation

Saline

Formol ether

Sat

saline

Zinc sulphate

Sheather

s sugar

STOOL EXAMINATION

Floatation concentration

Sat saline

Zn sulphate

Sheather

s sugar



Cestode eggs (non op)


Nematode eggs?????


Hookworms???????


Trichostong
؟؟؟؟؟؟؟؟؟؟؟


Egg of S.m.


Eggs of small tapeworms


Cysts



Crypto, Iso. oocysts

Tin container

20
min

Centrif.
2
min

Seive

Clean light eggs &
cysts

STOOL EXAMINATION

MACROSCOPIC

MICROSCOPIC

OTHERS


Consistency


Colour


Composition


Culture


Cellophane tape


Baeremann tech.


Ova quantitaion


(Stoll & Kato)


Temprory

Permanent

Diect saline smear

Iodine smear

Concentration techniques

Sedimentation

Floatation

Saline

Formol ether

Sat

saline

Zinc sulphate

Sheather

s sugar

STOOL EXAMINATION

Permanent Stained smears


Iron haematoxylin stain


Trichrome stain



Modified Ziehl Neelsen stain (Crypto.)

STOOL EXAMINATION

MACROSCOPIC

MICROSCOPIC

OTHERS


Consistency


Colour


Composition



Cellophane tape



Culture


Baeremann tech.


Ova quantitaion


(Stoll & Kato)


Temprory

Permanent

Diect saline smear

Iodine smear

Concentration techniques

Sedimentation

Floatation

Saline

Formol ether

Sat

saline

Zinc sulphate

Sheather

s sugar

STOOL EXAMINATION

Kato technique

Mesh screen

Template

Hole

Remove the template

Cellophane soaked by glycerin
(clears faeces
(

Egg count/ g stool


Egg quant. Of: Ascaris, T. trich., Hookworms, S. mansoni

STOOL EXAMINATION

Stoll

s technique

NaOH


4
g Stool

Erlynmeyer flask

56
CC

60
CC

Shake well

0.15
CC

Egg count/ slide

Eggs/
1
g= Eggs/slideX
100

Egg/day=Eggs/
1
g X stool wt/g in
24
hrs

24
hr stool


Egg quant. Of: Ascaris, T. trich., Hookworms, S. mansoni

STOOL EXAMINATION

Baermann

s technique

Warm

water

Stool/soil

seive

Glass funnel

clamp

30
min

25
-
50

CC


centrifuge

Detec. Of Nematode L. /stool, soil

STOOL EXAMINATION



Cultures for Nematode larvae

Filter paper culture

Scanty infection

Larvae of:



St. stercoralis (A,L)



Hookworms



Trichostrong

Water

Sealed petri dish

Filter paper

Slide

DIAGNOSIS

DIRECT

INDIRECT

MOLECULAR

Urine

Stool

Sputum

Biopsy

Aspirates

Blood

PCR

DNA probes

IHAT

LAT

IFAT

ELISA

CFT

DEIDT

SPUTUM EXAMINATION

MACROSCOPIC

MICROSCOPIC

Appearance

Concentration

Bloody (Parag)

Rusty brown (Parag)

NaOH

Sputum

Centfifuge

Parasites/sputum

P living in lung

P migrating in lung

P resulting from rupture of



P. westermani eggs



St. stercoralis



Ascaris



Hookworm (filariform L)



Hydatid cyst (sand)



Amoebic abcess (troph)

BIOPSY SPECIMEN

SKIN SNIP

MUSCLE BIOPSY

RECTAL BIOPSY

O. Volvulus mf

T. Spiralis larvae

Schistosoma egg



Raise skin by needle



Slice by scissors



Put snip in normal saline



Examine

Muscle digestion with HCl + pepsin

ASPIRATES EXAMINATION

CSF

Duodenal aspirates

BM aspirates

Cutanoeus ucler



Afr. Tryp. (trypom)



FLA (troph)



M.f. of loa loa



L. of T spiralis




G. lamb troph



Crypto oocyst



St sterc. Rh L.



Fasciola eggs



L. donovani a,ast.


T. cruzi amast.


P. falciparum.


Leishmaniasis

D. intubation

D capsule (Enterotest)

Direct stain (amast)

NNN (promast)


Lumbar puncutre



Centrifuge


Examine sed.

floor

Edge

DIAGNOSIS

DIRECT

INDIRECT

MOLECULAR

Urine

Stool

Sputum

Biopsy

Aspirates

Blood

PCR

DNA probes

IHAT

LAT

IFAT

ELISA

CFT

DEIDT

BLOOD EXAMINATION

Blood films

Buffy coat films

QBC technique

Knott

s conc. tech.

Thin

Thick

BLOOD EXAMINATION

BLOOD FILMS


Thin


Thick

Bld drop

spread

Air dry

methyl alcohol

Geimsa

Air dry

Geimsa

Circular motion

Malaria, Babesia, Filaria, Tryp.

BLOOD EXAMINATION

Buffy coat film


centrifuge

RBC

WBC (BC)

plasma

Citrated bld

30
min

Air dry

Fix

spread

Geimsa

Tryp., L. donovani


BLOOD EXAMINATION

QBC technique


centrifuge

RBC

RBC +parasite

Microhaematocrit tube

Acridine orange

Malaria, Filaria, Trypanosomes

BLOOD EXAMINATION

KNOTT

S CONC. TECHNIQUE


10
ml

1
ml

Air dry

fix

Geimsa


Citrated bld

Formalin
2
%

sediment

2
min

centrifuge

Filaria

INDIRECT IMMUNOLOGICAL
METHODS


Scanty infection.


Tissue parasite no portal of exit (Hydatid dis.)


Migratory stage (Fasciola)


Chronic infection fibrosis (Bilharziasis)


INDIRECT IMMUNOLOGICAL METHODS

Antigen detection

Antibody detection



More specific



More accurate.



Active infection



Early



Quantitative



Ab remain in serum for

months even after cure

INDIRECT IMMUNOLOGICAL METHODS

IHAT

LAT


+

Sensitized

Sheep

s RBC

(O

ve)

Ag

Patient

s serum

(?? AB)

Agglutination

+

Agglutination

Ag

Latex particle

Patient

s serum

(?? AB)

INDIRECT IMMUNOLOGICAL METHODS

INDIRECT FLUORESCENT ANTIBODY TEST



parasite

Patient

s serum

(?? AB)

Anti human AB

fluorescein

INDIRECT IMMUNOLOGICAL METHODS

ELISA



OPD

OPD

Flat bottom plastic micrititre plate

Ag

Patient

s serum

(?? AB)

Anti human AB

Peroxidase E

AB

INDIRECT IMMUNOLOGICAL METHODS

CFT



Ag

Patient

s serum

(?? AB)

complement

Anti sheep AB

Sheep

s RBC

Tube / microplate

AB

INDIRECT IMMUNOLOGICAL METHODS

Double Electro Immuno Diffusion

Ag

Ab

Buffered gel

Electric current

Line of ppt

INDIRECT IMMUNOLOGICAL METHODS

Immunodiagnostic Strip Test (Dip Stick Test) Ag

Nitrocellulose strip

Monoclonal Ab

Coloured dye

Pt bld (?Ag)

Malaria, Filaria, African tryp.

DIAGNOSIS

DIRECT

INDIRECT

MOLECULAR

Urine

Stool

Sputum

Biopsy

Blood

Aspirates

PCR

DNA probes

IHAT

LAT

IFAT

ELISA

CFT

DEIDT

MOLECULAR BIOLOGICAL TECHNIQUES

DNA Probes


DNA Probe

Commercially prepared DNA sequence

Radio active material

Nitrocellulose paper

Sample (Serum/ stool)

?? parasite

+ve parasite

Hybridization

Radioactivity

MOLECULAR BIOLOGICAL TECHNIQUES


Polymerase Chain Reaction (PCR)

Single stranded DNA




Replication








Detection

T cruzi, T gondii