lab oratory diagnosis of parasitic diseases

LAB ORATORY LAB ORATORY DIAGNOSIS OF DIAGNOSIS OF

PARASITIC DISEASESPARASITIC DISEASES

DEPARTEMENT OFDEPARTEMENT OF PARASITOLOG PARASITOLOGYY

FAFACCULTULTY OF MEDICINEY OF MEDICINE

UNIVERSITAS PADJADJARANUNIVERSITAS PADJADJARAN

DEPARTEMENT OFDEPARTEMENT OF PARASITOLOG PARASITOLOGYY

FAFACCULTULTY OF MEDICINEY OF MEDICINE

UNIVERSITAS PADJADJARANUNIVERSITAS PADJADJARAN

INTRODUCTION

DIAGNOSIS FOR PARASITIC INFECTION

Anamnesa – ( The history should include details of the presenting complaint )

Physical examination Laboratory examination espectially

Parasitological diagnosis Imunodiagnosis

DIAGNOSIS FOR PARASITIC INFECTION

Anamnesa – ( The history should include details of the presenting complaint )

Physical examination Laboratory examination espectially

Parasitological diagnosis Imunodiagnosis

AN

PHYSICAL EXAMINATIONPHYSICAL EXAMINATION

ANAMNESIS ANAMNESIS

ASPESIFICASPESIFIC

CLINICAL FEATURES (Symptom and Sign)

CLINICAL FEATURES (Symptom and Sign)

SPESIFICSPESIFIC

LABORATORY EXAMINATIONLABORATORY EXAMINATION

1

3

2

INTRODUCTION

-Fever

-Gastrointestinal symptoms

-Fever with respiratory system

-Neurological symptoms

-Fever and meningitis / Encephalitis

-Cutaneous symptoms

-Fever

-Gastrointestinal symptoms

-Fever with respiratory system

-Neurological symptoms

-Fever and meningitis / Encephalitis

-Cutaneous symptoms

INTRODUCTION

DEPEND ON : Habitat ParasiteParasite distribution

DEPEND ON : Habitat ParasiteParasite distribution

•Very important for examination Helminth parasite and Protozoa parasite

• Repeating of Laboratory examination occasionally be needed

*Should be understood about examination technique,morphology of

parasite and life cycle of parasite

•Very important for examination Helminth parasite and Protozoa parasite

• Repeating of Laboratory examination occasionally be needed

*Should be understood about examination technique,morphology of

parasite and life cycle of parasite

TYPE OF SPECIMEN :1. Stool 2. Blood3 Urine4 Sputum5. Vaginal discharge, urethral

discharge6. Skin scrapings7. Liquor Cerebro Spinal 8. Tissue biopsy9. Nasal discharge10. Corneal scrapings

TYPE OF SPECIMEN :1. Stool 2. Blood3 Urine4 Sputum5. Vaginal discharge, urethral

discharge6. Skin scrapings7. Liquor Cerebro Spinal 8. Tissue biopsy9. Nasal discharge10. Corneal scrapings

LABORATORY EXAMINATIONLABORATORY EXAMINATION3

IMUNODIAGNOSISIMUNODIAGNOSIS4

IMPORTANTIMPORTANT

INTRODUCTION

FECAL SPECIMENSFECAL SPECIMENS

Immediatelly have to examine :

Liquid specimens should be examined within 30 minute of passage

Soft (semiformed) specimens 1 hour

Formed specimens 24 hour after passage

If this time is not possible, the specimen should be placed in one of the available fixatives :

Formalin 10%, MIF (Merthiolate Iodine Formalin), PVA (Polyvinyl Alcohol)

Generally Helminthic eggs more endure without preservation than intestinal protozoa

Immediatelly have to examine :

Liquid specimens should be examined within 30 minute of passage

Soft (semiformed) specimens 1 hour

Formed specimens 24 hour after passage

If this time is not possible, the specimen should be placed in one of the available fixatives :

Formalin 10%, MIF (Merthiolate Iodine Formalin), PVA (Polyvinyl Alcohol)

Generally Helminthic eggs more endure without preservation than intestinal protozoa

INTRODUCTION


The specimens should not be contaminated with

Water – because water may contain free-

living organisms that can be mistaken for

human parasites

Urine – may destroy motile organisms

Prior to examination , fecal specimens should

never be incubated or frozen.

A chatartic with an oil base should not be used,

and a stool softener (taken either orally or as a

suppository) is usually inadequate for obtaining a

purged specimen.

The specimens should not be contaminated with

Water – because water may contain free-

living organisms that can be mistaken for

human parasites

Urine – may destroy motile organisms

Prior to examination , fecal specimens should

never be incubated or frozen.

A chatartic with an oil base should not be used,

and a stool softener (taken either orally or as a

suppository) is usually inadequate for obtaining a

purged specimen.

INTRODUCTION


Repeating fecal examination after therapy :

Ascariasis, 2-3 weeks after therapy

Protozoa infection, 3-4 weeks after

therapy

Taeniasis, 5-6 weeks after therapy

Repeating fecal examination after therapy :

Ascariasis, 2-3 weeks after therapy

Protozoa infection, 3-4 weeks after

therapy

Taeniasis, 5-6 weeks after therapy

EXAMINATION OF HELMINTH PARASITE

SPECIMENS( most important )

SPECIMENS( most important )

FECAL SPECIMENS FECAL SPECIMENS

BLOOD AND TISSUE SPECIMENS BLOOD AND TISSUE SPECIMENS

&&


FECAL SPECIMENS FECAL SPECIMENS ?

For examination of helminth egg Most important for examination intestinal

nematode including “Soil Transmitted Helminths” :

Ascaris lumbricoides Trichuris trichiura Hook worm : - Necator americanus

and Ancylostoma duodenale Strongyloides stercoralis

For examination of helminth egg Most important for examination intestinal

nematode including “Soil Transmitted Helminths” :

Ascaris lumbricoides Trichuris trichiura Hook worm : - Necator americanus

and Ancylostoma duodenale Strongyloides stercoralis


FECAL SPECIMENS FECAL SPECIMENS

DIRECT WET SMEAR

QUALITATIVE EXAMINATIONQUALITATIVE EXAMINATION

ANOTHER QUALITATIVE EXAMINATION AND

QUANTITATIVE EXAMINATION

TO BE STUDIED IN FECAL EXAMINATION

LABORATORY TECHNIQUE FOR EXAMINATION OF HELMINTH PARASITE

LABORATORY TECHNIQUE FOR EXAMINATION OF HELMINTH PARASITE

LABORATORY TECHNIQUE FOR

EXAMINATION HELMINTH PARASITE

INTESTINAL

HELMINTH

BLOOD AND

TISSUE

HELMINTH

FECAL SPECIMENS

BLOOD AND TISSUE

HELMINTH

Click “Esc” buttonWhen finished

MACROSCOPIC

EXAMINATION

MICROSCOPIC

EXAMINATION

EXAMINATION TECHNIQUE FORHELMINTH PARASITE IN FAECES

LABORATORY TECHNIQUE FOR EXAMINATION OF INTESTINAL

HELMINTH

QUALITATIVE

QUANTITATIVE

*Consistency (hard,formed,soft,diarrhea)

*Colour

*Odor

*Foreign bodies (blood, mucus,parasite)


MICROSCOPIC

QUALITATIVE

STOLL DILLUTION

METHOD

KATO – KATZ

CELLOPHANE THICK

SMEAR METHOD

QUANTITATIVE

EXAMINATION TECHNIQUE OF INTESTINAL HELMINTH


DIRECT WET SMEAR METHOD

FLOTATION METHOD

MODIFICATION MERTHIOLATE IODINE

FORMALDEHYDE (MIF)

CELLOTAPE TAPE METHOD

FORMALDEHYDE ETHER SEDIMENTATION

(RITCHIE’S METHOD)

METODA KATO

Fast Severe infection Reagens

NaCl Physiologis (0,9%), or Eosin 2%

DIRECTDIRECT WET SMEAR METHOD WET SMEAR METHOD

MIMICCROSROSCCOPIOPICCQQUALITATIUALITATIVEVE


DIRECT WET SMEAR METHODDIRECT WET SMEAR METHOD


1-2 dropsNaCl 0,9%

oreosin 2%

Place 2 mg faeceson microscope slide

Emulsify 2 mg faeces in NaCl 0.9% dropPlace 1-2 drops NaCl 0,9% or Eosin

2% on microscope slide


Place 2 mg faeces on microscope slide


SALINE SALINE WET SMEAR METHODWET SMEAR METHODPlace 2 mg faeces

on microscope slide

Emulsify 2 mg faeces in NaCl 0,9% drop

Place coverslip overthe suspension

1-2 dropsNaCl 0,9%

Emulsify 2mg faeces in the saline dropPlace coverslip over suspension

Examine using the lower power objective 10x or 40x


FLOATATION METHODFLOATATION METHOD

Berdasarkan BJ telur < BJ larutan

Berguna untuk infeksi ringan Larutan yang dipergunakan :

NaCl jenuh, atau Gula jenuh

FLOTATION METHODWITH CENTRIFUGATION

WITHOUT CENTRIFUGATION



NaCl saturatedfaeces

stirringglass

Ose

20 minutes

10 gr faeces mixed with 200 ml NaCl saturated until homogenous solution

Take down 1-2 drops of Surface layer with Ose andPlace on microscope slide

10 gr. faeces + 200 cc NaCl saturated

Place coverslip overthe solution

FLOATATION METHODFLOATATION METHODWITHOUT CENTRIFUGATION

Take down 1-2 drops the surface layer with OsePlace coverslip over microcope slide

Examine under low power objective



Stirringglass

filtered

Centrifuge 100 rpmfor 5 minutes

Transfer 10 gr faeces and emulsify in 200 ml NaCl saturated until

homogenous solution10 gr. faeces + 200 cc NaCl saturated

FLOATATION METHODFLOATATION METHODWITH CENTRIFUGATION



stirringglass

filtered

centrifuge 100 x/mnt5 minutes

Filtered with tea filter10 gr. faeces

+ 200 cc NaCl saturated



stirringglass

filtered

centrifuge 100 rpmFor 5 minutes

Decant into centrifuge tube





Stirringglass

filtered

centrifuge 100 rpmFor 5 minutes

Centrifuge 100 rpm for 5 minutes

10 gr.faeces + 200 cc NaCl saturated




stirringglass

filtered

Centrifuge 100 rpmFor 5 minutes

Take down solution from surface layer with Ose




Place solution on microscope slide and Place coverslip over the microscope slide.

Examine under low power objective 10 x or 40 x.

Place the solution onmicroscope slide Place coverslip over

microscope slide



For identification eggs from intestinal helminth, Ameba and Giardia lamblia

Solution 1 : 250 ml aquadest 200 ml thimerosal (1:1.000) 25 ml formalin 5 ml glycerin

Solution 2 : Fresh lugol solution5%

MERTIOLAT IODIN FORMALIN MODIFICATION MERTIOLAT IODIN FORMALIN MODIFICATION (MIF MODIFICATION)(MIF MODIFICATION)


MERTMERTHHIOLAT IODIN FORMALIN MODIFIIOLAT IODIN FORMALIN MODIFICATIONCATION (MIF MODIFI(MIF MODIFICATIONCATION ) )

It’s good for staining and conservation of cyst of intestine protozoa and worm egg


(+) 7 ml. ether(40 C)


Gelaspengaduk

5 ml base solution 10,5 ml base solution 2

0,5 gr faeces

filtered Shake until homogenous

centrifuge1.000-3.000 x/mnt1 minutes

shake until homogenous


Add 7 ml ether (40 C)

Open the tube, let it 2 minutes, centrifuge for 1 minutes (1.000-3.000 rpm)


Throw away the supernatan,take sediment with pipette

Place the sediment onmicroscope slide

Place coverslip overmicroscope slide


Examine under low power objective

CELLOTAPE METHODCELLOTAPE METHOD

The egg adhere on perianal area, so rarely found in

faeces (5 %). To find this parasite we need Scotch

Adhesive tape Swab from Graham or Cellotape

Method



To examine the egg of Enterobius vermicularis Children 1-10 years old Doing in the morning before take a bath or wash

the anus with water after defecating Transparent plastic plaster (2 x 1,5 cm)patched

to skin around the anus Press the plaster, then lift slowly Patched to the object glass, examine under the

microscope


Concentration MethodConcentration Method

practical, simple, for ova examination in stool : 1 gr faeces, put into the reaction tube, add aquadest,

mixed until homogenous, put imyo the centrifuge tube Centrifuge with velocity 3.000 rpm for 1 mnt Throw away the supernatan, take the sediment with

pipette Place the sediment on microscope slide, place

coverslip on microscope slide


Put in faeces solution to the centrifuge tube

Centrifuge 3.000 x/mnt,1 minutes


Shake until homogenous

1 gr faeces

Stirring glass

Aquadest


Throw away supernatan, take sediment with pipette

Place sediment on microscope slide



place coverslip on microscope slide

Examine under the microscope

CELLOPHANE THICK SMEAR METHOD (KATO METHOD)(KATO METHOD)

Practical, simple, and cheap Can be used in mass examination Examination needs more stool, so the eggs can be found

much more Morphology of egg is clear


CELLOPHANE THICK SMEAR METHOD

(KATO METHOD(KATO METHOD))

Substance : Solution

100 ml aquadest/fenol 6% 100 ml glycerin (supaya kotoran tinja jadi jernih) 1 ml malachit green solution (supaya mata tidak

silau)

Cellophane tape, 2,5 x 3 cm, soak in solution for >24 hours


CELLOPHANE THICK SMEAR METHOD(KATO METHOD)(KATO METHOD)

Technique : Take 20-50 mg faeces ( as large as red bean ) Put on object glass, spread out Cover with cellophane, pressing the faeces until

flat and spread out under the cellophane Drain the excessive fluid with filter paper Let it 20-30 minutes Examine under the microscope


CELLOPHANE THICK SMEAR METHOD(KATO METHOD)(KATO METHOD)

100 part. Aquadest/fenol 6%100 part. Glycerin1 part Malachit green solution

Cut the cellophane2,5 x 3 cm

Soaked

> 24 hours

20-50 grfaeces

Soaked cellophane

faeces


Solution used : NaOH 0,1 N (faeces solvent) or KOH 10%

Good for severe and moderate infection

Less good for mild infection

STOLL METHODSTOLL METHOD

MIMICCROSROSCCOPIOPICCQQUANTITATIUANTITATIVEVE

MICROSCOPICMICROSCOPICQUANTITATIVEQUANTITATIVE

56 m l60 m l

56 m l60 m l

Butir G elas

Shake

Let it 1 night

or3-4 hours but shake it

longer

56 m l60 m l

faeces

NaOH solution 0,1N


56 ml 60 ml


Shake and take 0,15 ml

56 m l60 m l



Formula :

amount of egg in 1 gram feces = amount of seen egg (miroscopic) x 100



ENUMARATION NaOH = 56 ml, feces 4 ml ~ 4gr 4 gr feces in 60 ml or 1 gr feces in 15 ml In the 0.15 ml of stool solution,

can be found y eggs So, in the 15 ml, is found y x 100

eggs(~ 1 gr stool)



Infection level based on amount of egg in each gram feces and amount of helminth

MILD MODERATE SEVERE

< 7.000 7.000-35.000 > 35.000

A. lumbricoides

5 or less 6-25 > 25


A. duodenale

< 3.000 3.000-10.000 > 10.000

20 or less 21-100 > 100


N. americanus

< 2.000 2.000-7.000 > 7.000

50or less 51-200 > 200

AMOUNT OF HELMINTH

AMOUNT OF EGG PER-GRAM

FAECES

INFECTION LEVEL

Infectid by

SOURCE : PARASITIC DISEASES PROGRAME. WHO. GENEVA, 1981


KATO-KATZ METHODKATO-KATZ METHOD


Tolls : Object glass cellotape, thick 40 , size 3 x 3 cm Thick carton, make hole with fixed

volume to print faeces as weight as 30 mg

Palm leaf rib, oily papper, wire netting



Malachite-Green solution : 100 ml aquadest 100 ml glycerin (in order to make the

feces clear) 1 ml malachite green solution

Soak the cellotape in solution for > 24 hours



FILTERING FAECES

Put 5 gr faeces on the oily papper, put wire netting on it then press

Put the holed carton on the object glass, print the filtered faeces on holed carton

Lift the carton Cover the faeces

with soaked cellotape

Press, spread out Let it 30 mnt Examine under the

microscope

PRINTING FAECES MAKING PREPARAT



Filtering faeces

Oily papper

Wire netting

press

faeces (5 gr)

TEKAN

Wire netting

faeces (5 gr)

Object glass

Holed carton

Filtered faeces

printed

Printed faeces Object glass

Soaked cellotape

Printing faeces Making THE SPECIMEN

EXAMINE UNDERTHE MICROSCOPE



WHO (1981) EGG PRODUCTION PER-DAY :

A. lumbricoides : 200.000 A. doudenale : 10.000-25.000 N. americanus : 5.000-10.000

WEIGHT OF FAECES PER-DAY Children : 70 gram Adult : 2 x children

FORMOL ETHER SEDIMENTATION METHODFORMOL ETHER SEDIMENTATION METHOD(RITCHIE)

MICROSCOPICMICROSCOPICQUALITATIVEQUALITATIVE

(0,5 ml faeces) + (1-2 ml aquadest), shake, + (10-12 ml aquadest), shake

Filter Centrifuge 1.000 rpm, 1 mnt + (1 ml formalin 10%) + (formalin 10% sampai

volume 8 ml), let it 10 mnt + (3ml ether), shake 15-20 second Centrifuge 2.000 rpm, 1-2 mnt Throw away the supernatan, place the

sediment on microscope slide, examine under the microscope


shake

0,5 ml. faeces

1-2 ml. aquadest

shake

10-12 ml. aquadest

filter

centrifuge1.000 rpm, 1 minute


centrifuge 1.000 rpm, 1 minute


+ (1 ml formalin 10%) + (formalin 10% until volume 8 ml), let it 10 minutes

LET IT(10 minutes)

3 ml. ether

1 ml. Formalin 10%+ Formalin 10% until

volume 8 ml

SHAKE(15-20 seconds)

CENTRIFUGE2.000 rpm, 1-2 minutes


Throw away the supernatan, place the sediment on microscope slide, examine

under the microscope

USING FORMALIN SOLUTION 3,5 % OR 4%USING FORMALIN SOLUTION 3,5 % OR 4%

CONSERVATION AND STORAGE METHODCONSERVATION AND STORAGE METHODEGG AND ADULT WORM IN FAECESEGG AND ADULT WORM IN FAECES

Making formalin solution 3,5 % Or 4% : 1 part of formalin solution 35% or 40% mixed with 9

part of running water, then put into closed bottle

Put the faeces into the bottle and closed tightly For organ that attacked by parasite, wash cleanly

before put into the bottle

Laboratory tecknique for examination of Filaria sp.Laboratory tecknique for examination of Filaria sp.

BLOOD AND TISSUE NEMATODEBLOOD AND TISSUE NEMATODE((Blood specimensBlood specimens))

DIRECT METHOD Blood taken of from finger tip Thin blood smear

Qualitative Determine microfilariae in peripheral blood

vessels Not for species identification

Thick blood smear Quantitative Measured blood releasefrom finger tip with

mikropipet (160 m), make thick blood smear. CONCENTRATION METHOD

Take blood vein More sensitive than direct method

Click “esc” buttonWhen finished

SUBJECT MATERIAL SUBJECT MATERIAL � HOW TO EXAMINE AND INTERPRET INTESTINAL PROTOZOA

- Direct wet mount - Modified Merthiolate-Iodine -Formalin (MIF)

� HOW TO EXAMINE AND INTERPRET BLOOD PROTOZOA - Thin Blood smear with Giemsa staining - Thick Blood smear with Giemsa staining

� EXAMINATION FOR Trichomonas vaginalis� HOW TO PREPARE PERMANENT (FIXED) MOUNT� - Blood parasites

- Trichomonas vaginalis- Ameba, using Heidenhein method and staining

� METHODS FOR PREPARING COLOR STAININGS

� HOW TO EXAMINE AND INTERPRET INTESTINAL PROTOZOA - Direct wet mount - Modified Merthiolate-Iodine -Formalin (MIF)

� HOW TO EXAMINE AND INTERPRET BLOOD PROTOZOA - Thin Blood smear with Giemsa staining - Thick Blood smear with Giemsa staining

� EXAMINATION FOR Trichomonas vaginalis� HOW TO PREPARE PERMANENT (FIXED) MOUNT� - Blood parasites

- Trichomonas vaginalis- Ameba, using Heidenhein method and staining

� METHODS FOR PREPARING COLOR STAININGS

(1). Direct wet mount exam (1). Direct wet mount exam

Purpose :

For quick and simple examination

Purpose :

For quick and simple examination

LAB METHODS FOR INTESTINAL PROTOZOA LAB METHODS FOR INTESTINAL PROTOZOA LAB METHODS FOR INTESTINAL PROTOZOA LAB METHODS FOR INTESTINAL PROTOZOA

- For trophozoite form of ameba, use 2% eosin solution- For trophozoite form of ameba, use 2% eosin solution

- For cyst and nucleus of amoeba use lugol solution

(2% of Iodine + 3% potasssium Iodine)- For cyst and nucleus of amoeba use lugol solution

(2% of Iodine + 3% potasssium Iodine)

Using a small stick, add small amount of feces on top of an object glass

Add few drops of physiological saline solution NaCl/lugol/eosin 2% on top

PreparationPreparation : :

Distribute/spread evenly and cover with coverglass

Examine under low power microscope

LAB METHODS FOR INTESTINAL PROTOZOALAB METHODS FOR INTESTINAL PROTOZOA LAB METHODS FOR INTESTINAL PROTOZOALAB METHODS FOR INTESTINAL PROTOZOA

(1). (1). Direct wet mount examination Direct wet mount examination

(2). Modified Method Merthiolate-Iodine-Formaline (MIF)

PurposePurpose : :Good for diagnosis of Ameba and Giardia in feces

LAB METHOD FOR INTESTINAL PROTOZOA LAB METHOD FOR INTESTINAL PROTOZOA

Click “Esc”buttonWhen finished

Ingredients used Ingredients used ::Basic solution (1) :

–250 ml aquadest

–200 ml tincture of merthiolate

–25 ml formaldehyde

Basic solution (2) :

–lugol 5 % (not to be kept > 3 weeks)

Both basic solutions should be kept in brown colored bottle

Modified Method Merthiolate-Iodine-Formaline (MIF)

LAB METHOD FOR INTESTINAL PROTOZOA LAB METHOD FOR INTESTINAL PROTOZOA

Click “Esc”buttonWhen finished

Acidic in character Foul smelling Produce less mucus compared to

bacillary dysentery, less sticky With or without blood (blood may be

found in solid feces) In some cases mucosal wall may come

out

Acidic in character Foul smelling Produce less mucus compared to

bacillary dysentery, less sticky With or without blood (blood may be

found in solid feces) In some cases mucosal wall may come

out

CHARACTERISTIC OF FECES WITH AMEBA CHARACTERISTIC OF FECES WITH AMEBA

MACROSCOPIC MACROSCOPIC


MACROSCOPICMACROSCOPIC

Source :A Colour Atlas of Clinical Parasitology. Tomio Yamaguchi. Alih Bahasa : Lesmana Padmasutra, dkk.

Feces of amebic dysenteric patient, plenty of trophozoite stage

CHARACTERISTICS OF FECES WITH AMEBACHARACTERISTICS OF FECES WITH AMEBA

MACROSCOPICMACROSCOPIC

Source : Color Atlas of Medicine and Parasitology. 1977. W. Peters & H.M. Gillers

Feces of amebic dysenteric patient, loose, slimy and bloody feces

CHARACTERISTICS OF FECES WITH AMEBACHARACTERISTICS OF FECES WITH AMEBA

MICROSCOPICMICROSCOPIC Plenty of bacteria Entamoeba histolytica (+) containing erythrocytes Erythrocytes in reuleaux (chain) formation Charcot-Leyden crystals (unspecific for ameba dysentery, can

be found also in other parasitic infection e.g. Strongyloides stercoralis) – from eosinophil desintegration

Pus less abundant compared to bacillary dysentery, if no secondary infection

Macrophages in bacillary dysentery mimics Entamoeba histolytica but nucleus and pseudopods differs

Cyst found in carrier patients and cases with light infection

Plenty of bacteria Entamoeba histolytica (+) containing erythrocytes Erythrocytes in reuleaux (chain) formation Charcot-Leyden crystals (unspecific for ameba dysentery, can

be found also in other parasitic infection e.g. Strongyloides stercoralis) – from eosinophil desintegration

Pus less abundant compared to bacillary dysentery, if no secondary infection

Macrophages in bacillary dysentery mimics Entamoeba histolytica but nucleus and pseudopods differs

Cyst found in carrier patients and cases with light infection


microscopicmicroscopic

Charcot-Leyden crystals(From eosinophil desintegration)

Source;Color Atlas of Medical Parasitology. Prayong Radomyos, dkk.

Blood slides with Giemsa staining

PurposePurpose : :For the examination of blood protozoa, e.g. :

Plasmodium,

Trypanosoma, Babesia etc.

Prepared in two stagesPrepared in two stages : :(1). Prepare the blood smear

(2). Do the color staining

EXAMINATION METHODS FOR BLOOD PROTOZOA EXAMINATION METHODS FOR BLOOD PROTOZOA

Place blood slide in upright position, allow to dry, keep away from dust or small insects.

EXAMINATION METHOD FOR BLOOD PROTOZOA EXAMINATION METHOD FOR BLOOD PROTOZOA


Preparation of thin blood film

EXAMINATION METHOD FOR BLOODEXAMINATION METHOD FOR BLOOD PROTOZOA PROTOZOA


Staining procedure

Fix with methyl alcohol (3-5) minutes

Stain with standard Giemsa for 45 minutes

Wash slowly with tap water

ALLOW TO DRY

EXAMINATION METHOD FOR BLOODEXAMINATION METHOD FOR BLOOD PROTOZOAPROTOZOA

Thick blood smear with Giemsa stain

Purpose :Rapid examination of blood protozoa

(for mass survey and screening )

Conducted in two stages :(1). Prepare thick blood film(2). Staining the blood film


(1). Preparation of thick blood smear


Blood is prepared similar to thin blood smear Place 1-2 drops of blood on a glass slide Spread evenly, forming a circle of 1 - 1,5 cm in diameter Allow to dry, keep away from dust or insect


(2). Staining procedure

ALLOW TO DRY

Stain with standard Giemsa for 45 minutes

Rinse slowly with tap water


NO FIXATION !!!

� THIN BLOOD SMEAR– Morphology and stages of Plasmodium clearly defined– Erythrocytes are intact (due to fixation)– Slow to prepare – Used for examination of moderate and heavy infection

� THICK BLOOD SMEAR – Morphology and stages of Plasmodium not clearly defined– Erythrocytes are lysed, only stroma from erythrocytes are seen – Preparation and staining are faster (can be used for mass survey

examination)– Commonly used for light infection

COMPARISON OF THIN AND THICK BLOOD SMEAR


EXAMINATION OF Trichomonas vaginalisEXAMINATION OF Trichomonas vaginalis

Methods of examination :(1). Direct examination

(2). Culture

Methods of examination :(1). Direct examination

(2). Culture

Direct method :(1). Direct wet mount

(2). Staining

EXAMINATION OF Trichomonas vaginalisEXAMINATION OF Trichomonas vaginalis

Sample for examination :(1). Women : vaginal, or urethral discharge

(using vaginal spiculum)(2). Men : discharge from urethra or prostate,

and centrifugated urine sample

DIRECT EXAMINATION

Materials used :-Test tube containing 5% glucose (in physiological saline solution)

- Cotton bulb

Cotton bulb

Glucose 5%

Examination for Trichomonas vaginalisExamination for Trichomonas vaginalis

Terima kasih ………………….

April2005

lab oratory diagnosis of parasitic diseases

Documents

examination helminth

hourformed specimens

liquid specimens

protozoa parasite

important fecal specimensblood

morphology of parasite

purged specimen

therapy protozoa infection