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laboratory diagnosis

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LABORATORY DIAGNOSIS OF PARASITIC INFECTIONS : 

LABORATORY DIAGNOSIS OF PARASITIC INFECTIONS By: Dr.Maha jaafar

Case diagnosis : 

Case diagnosis History (Age, occupation, residency, previous infection) Complaint Clinical examination Invesigations - Laboratory investigations - Radiology - Surgical intervention (Exploratory) Provisional diagnosis Confirm the diagnosis

Slide 4: 

Acetic acid RBC haemolysis Clear ova Ether Dissolve fat M.f

URINE EXAMINATION : 

URINE EXAMINATION SEDIMENTATION CONCENTRATION 15-20 min Centrifuge (2 min) Clean conical glass receptacle

URINE EXAMINATIONMembrane filtration technique : 

URINE EXAMINATIONMembrane filtration technique air 10 ml urine Nucleopore filter Eggs of Schistosoma + Saline

URINE EXAMINATION : 

URINE EXAMINATION

Slide 10: 

STOOL EXAMINATION

STOOL EXAMINATIONTemporary : 

STOOL EXAMINATIONTemporary Saline smear Iodine smear saline Iodine 1% Huge number of: Eggs Protozoal troph. Motility (Amoeb, flagellates) Huge number of: Cyst morphological details

STOOL EXAMINATIONScanty infectionConcentration techniques : 

STOOL EXAMINATIONScanty infectionConcentration 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 EXAMINATIONSaline sedimentation : 

STOOL EXAMINATIONSaline sedimentation 10 g stool Saline Mesh wire gauze Conical flask Sediment Emulsify

STOOL EXAMINATION Formol Ether Sed. Conc. : 

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 : 

STOOL EXAMINATION Tin container 20 min Centrif. 2 min Seive Clean light eggs & cysts

STOOL EXAMINATIONPermanent Stained smears : 

STOOL EXAMINATIONPermanent Stained smears Iron haematoxylin stain Trichrome stain Modified Ziehl Neelsen satin (Crpto.)

STOOL EXAMINATION Kato technique : 

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 : 

STOOL EXAMINATION Stoll’s technique NaOH 4 g Stool Erlynmeyer flask 56 CC 60 CC Shake well 0.15 CC Egg count/ slide Eggs/1g= Eggs/slideX100 Egg/day=Eggs/1g X stool wt/g in 24 hrs 24 hr stool Egg quant. Of: Ascaris, T. trich., Hookworms, S. mansoni

STOOL EXAMINATION Baermann’s technique : 

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 : 

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

Slide 27: 

NaOH Sputum Centfifuge

Slide 30: 

floor Edge

BLOOD EXAMINATIONBLOOD FILMS : 

BLOOD EXAMINATIONBLOOD FILMS Thin Thick Bld drop spread Air dry methyl alcohol Geimsa Air dry Geimsa Circular motion Malaria, Babesia, Filaria, Tryp.

BLOOD EXAMINATIONBuffy coat film : 

BLOOD EXAMINATIONBuffy coat film centrifuge RBC WBC (BC) plasma Citrated bld 30 min Air dry Fix spread Geimsa Tryp., L. donovani

BLOOD EXAMINATIONQBC technique : 

BLOOD EXAMINATIONQBC technique centrifuge RBC RBC +parasite Microhaematocrit tube Acridine orange Malaria, Filaria, Trypanosomes

BLOOD EXAMINATIONKNOTT’S CONC. TECHNIQUE : 

BLOOD EXAMINATIONKNOTT’S CONC. TECHNIQUE 10 ml 1 ml Air dry fix Geimsa Citrated bld Formalin 2 % sediment 2 min centrifuge Filaria

INDIRECT IMMUNOLOGICAL METHODS : 

INDIRECT IMMUNOLOGICAL METHODS Scanty infection. Tissue parasite no portal of exit (Hydatid dis.) Migratory stage (Fasciola) Chronic infection fibrosis (Bilharziasis)

INDIRECT IMMUNOLOGICAL METHODS : 

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 METHODSINDIRECT FLUORESCENT ANTIBODY TEST : 

INDIRECT IMMUNOLOGICAL METHODSINDIRECT FLUORESCENT ANTIBODY TEST parasite Patient’s serum (?? AB) Anti human AB fluorescein

INDIRECT IMMUNOLOGICAL METHODSELISA : 

INDIRECT IMMUNOLOGICAL METHODSELISA OPD OPD Flat bottom plastic micrititre plate Ag Patient’s serum (?? AB) Anti human AB Peroxidase E AB

INDIRECT IMMUNOLOGICAL METHODSCFT : 

INDIRECT IMMUNOLOGICAL METHODSCFT Ag Patient’s serum (?? AB) complement Anti sheep AB Sheep’s RBC -ve Ab +ve Ab haemolysis No Sheep RBChaemolysis Tube / microplate AB

INDIRECT IMMUNOLOGICAL METHODSDouble Electro Immuno Diffusion : 

INDIRECT IMMUNOLOGICAL METHODSDouble Electro Immuno Diffusion +ve -ve Ag Ab Buffered gel Electric current Line of ppt

INDIRECT IMMUNOLOGICAL METHODSImmunodiagnostic Strip Test (Dip Stick Test) Ag : 

INDIRECT IMMUNOLOGICAL METHODSImmunodiagnostic Strip Test (Dip Stick Test) Ag Nitrocellulose strip Monoclonal Ab Coloured dye Pt bld (?Ag) +ve -ve Malaria, Filaria, African tryp.

MOLECULAR BIOLOGICAL TECHNIQUESDNA Probes : 

MOLECULAR BIOLOGICAL TECHNIQUESDNA 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) : 

MOLECULAR BIOLOGICAL TECHNIQUES Polymerase Chain Reaction (PCR) Single stranded DNA Replication Detection T cruzi, T gondii

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