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LABORATORY DIAGNOSIS  OF  PARASITIC  INFECTIONS By: Dr.Maha jaafar
Case diagnosis ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Provisional diagnosis Confirm the diagnosis
DIAGNOSIS DIRECT INDIRECT MOLECULAR Urine Stool Sputum Biopsy Blood Aspirates PCR DNA probes IHAT LAT IFAT ELISA CFT DEIDT
Acetic acid  RBC haemolysis Clear ova Ether Dissolve fat M.f URINE EXAMINATION MACROSCOPIC MICROSCOPIC colour white smoky Sedimentation concentration Membrane filtration Chyluria Blood Filaria S. haematobium
URINE EXAMINATION ,[object Object],[object Object],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 ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
URINE EXAMINATION Egg viability Live eggs Dead eggs ,[object Object],[object Object],[object Object],[object Object],[object Object]
STOOL EXAMINATION MACROSCOPIC MICROSCOPIC OTHERS ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Temprory Permanent Diect saline smear Iodine smear Concentration techniques Sedimentation Floatation Saline Formol ether Sat  saline Zinc sulphate Sheather’s sugar
STOOL EXAMINATION MACROSCOPIC EXAMINATION COLOUR CONSISTENCY COMPOSITION Adult PARASITES Pale=Steatorrhea (G.l) ,[object Object],[object Object],[object Object],?? Blood ?? Mucus (dysentry) *Ascaris worm *E. vermicularis *T. saginata
STOOL EXAMINATION MACROSCOPIC MICROSCOPIC OTHERS ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Temprory Permanent Diect saline smear Iodine smear Concentration techniques Sedimentation Floatation Saline Formol ether Sat  saline Zinc sulphate Sheather’s sugar
STOOL EXAMINATION Temporary ,[object Object],[object Object],saline Iodine 1% ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
STOOL EXAMINATION Scanty infection Concentration techniques Sedimentation Floatation ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
STOOL EXAMINATION MACROSCOPIC MICROSCOPIC OTHERS ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],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 ,[object Object],[object Object],Conical flask centrif. tube
STOOL EXAMINATION MACROSCOPIC MICROSCOPIC OTHERS ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],Temprory Permanent Diect saline smear Iodine smear Concentration techniques Sedimentation Floatation Saline Formol ether Sat  saline Zinc sulphate Sheather’s sugar
STOOL EXAMINATION Tin container 20 min Centrif. 2 min Seive Clean light eggs & cysts Floatation concentration Sat saline Zn sulphate Sheather’s sugar ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
STOOL EXAMINATION MACROSCOPIC MICROSCOPIC OTHERS ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],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 ,[object Object],[object Object],[object Object]
STOOL EXAMINATION MACROSCOPIC MICROSCOPIC OTHERS ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],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/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 Warm   water Stool/soil seive Glass funnel clamp 30 min 25-50   CC ,[object Object],Detec. Of Nematode L. /stool, soil
STOOL EXAMINATION    Cultures for Nematode larvae ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],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
NaOH Sputum Centfifuge SPUTUM EXAMINATION MACROSCOPIC MICROSCOPIC Appearance Concentration Bloody (Parag) Rusty brown (Parag)
Parasites/sputum P living in lung P migrating in lung P resulting from rupture of ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object]
BIOPSY SPECIMEN SKIN SNIP MUSCLE BIOPSY RECTAL BIOPSY O. Volvulus mf T. Spiralis larvae Schistosoma egg ,[object Object],[object Object],[object Object],[object Object],Muscle digestion with HCl + pepsin
floor Edge ASPIRATES EXAMINATION CSF Duodenal aspirates BM aspirates Cutanoeus ucler ,[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],[object Object],D. intubation D capsule (Enterotest) Direct stain (amast) NNN (promast) ,[object Object],[object Object],[object Object]
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 ,[object Object],[object Object],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 ,[object Object],Formalin 2 % sediment 2 min centrifuge Filaria
INDIRECT IMMUNOLOGICAL METHODS ,[object Object],[object Object],[object Object],[object Object]
INDIRECT IMMUNOLOGICAL METHODS Antigen detection Antibody detection ,[object Object],[object Object],[object Object],[object Object],[object Object],Ab remain in serum for  months even after cure
INDIRECT IMMUNOLOGICAL METHODS ,[object Object],[object Object],+ 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 -ve  Ab +ve  Ab haemolysis No Sheep RBChaemolysis Tube / microplate AB
INDIRECT IMMUNOLOGICAL METHODS Double Electro Immuno Diffusion +ve -ve 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) +ve -ve 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

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Diagnosis

  • 1. LABORATORY DIAGNOSIS OF PARASITIC INFECTIONS By: Dr.Maha jaafar
  • 2.
  • 3. DIAGNOSIS DIRECT INDIRECT MOLECULAR Urine Stool Sputum Biopsy Blood Aspirates PCR DNA probes IHAT LAT IFAT ELISA CFT DEIDT
  • 4. Acetic acid RBC haemolysis Clear ova Ether Dissolve fat M.f URINE EXAMINATION MACROSCOPIC MICROSCOPIC colour white smoky Sedimentation concentration Membrane filtration Chyluria Blood Filaria S. haematobium
  • 5.
  • 6. URINE EXAMINATION Membrane filtration technique air 10 ml urine Nucleopore filter Eggs of Schistosoma + Saline
  • 7.
  • 8.
  • 9.
  • 10.
  • 11.
  • 12.
  • 13.
  • 14.
  • 15. STOOL EXAMINATION Saline sedimentation 10 g stool Saline Mesh wire gauze Conical flask Sediment Emulsify
  • 16.
  • 17.
  • 18.
  • 19.
  • 20.
  • 21.
  • 22. 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
  • 23. 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
  • 24.
  • 25.
  • 26. DIAGNOSIS DIRECT INDIRECT MOLECULAR Urine Stool Sputum Biopsy Aspirates Blood PCR DNA probes IHAT LAT IFAT ELISA CFT DEIDT
  • 27. NaOH Sputum Centfifuge SPUTUM EXAMINATION MACROSCOPIC MICROSCOPIC Appearance Concentration Bloody (Parag) Rusty brown (Parag)
  • 28.
  • 29.
  • 30.
  • 31. DIAGNOSIS DIRECT INDIRECT MOLECULAR Urine Stool Sputum Biopsy Aspirates Blood PCR DNA probes IHAT LAT IFAT ELISA CFT DEIDT
  • 32. BLOOD EXAMINATION Blood films Buffy coat films QBC technique Knott’s conc. tech. Thin Thick
  • 33.
  • 34. BLOOD EXAMINATION Buffy coat film centrifuge RBC WBC (BC) plasma Citrated bld 30 min Air dry Fix spread Geimsa Tryp., L. donovani
  • 35. BLOOD EXAMINATION QBC technique centrifuge RBC RBC +parasite Microhaematocrit tube Acridine orange Malaria, Filaria, Trypanosomes
  • 36.
  • 37.
  • 38.
  • 39.
  • 40. INDIRECT IMMUNOLOGICAL METHODS INDIRECT FLUORESCENT ANTIBODY TEST parasite Patient’s serum (?? AB) Anti human AB fluorescein
  • 41. INDIRECT IMMUNOLOGICAL METHODS ELISA OPD OPD Flat bottom plastic micrititre plate Ag Patient’s serum (?? AB) Anti human AB Peroxidase E AB
  • 42. INDIRECT IMMUNOLOGICAL METHODS CFT Ag Patient’s serum (?? AB) complement Anti sheep AB Sheep’s RBC -ve Ab +ve Ab haemolysis No Sheep RBChaemolysis Tube / microplate AB
  • 43. INDIRECT IMMUNOLOGICAL METHODS Double Electro Immuno Diffusion +ve -ve Ag Ab Buffered gel Electric current Line of ppt
  • 44. INDIRECT IMMUNOLOGICAL METHODS Immunodiagnostic Strip Test (Dip Stick Test) Ag Nitrocellulose strip Monoclonal Ab Coloured dye Pt bld (?Ag) +ve -ve Malaria, Filaria, African tryp.
  • 45. DIAGNOSIS DIRECT INDIRECT MOLECULAR Urine Stool Sputum Biopsy Blood Aspirates PCR DNA probes IHAT LAT IFAT ELISA CFT DEIDT
  • 46. MOLECULAR BIOLOGICAL TECHNIQUES DNA Probes DNA Probe Commercially prepared DNA sequence Radio active material Nitrocellulose paper Sample (Serum/ stool) ?? parasite +ve parasite Hybridization Radioactivity
  • 47. MOLECULAR BIOLOGICAL TECHNIQUES Polymerase Chain Reaction (PCR) Single stranded DNA Replication Detection T cruzi, T gondii