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PARASITIC INFECTIONS
             BASICS IN DIAGNOSIS
                  Dr.T.V.Rao MD




12/10/2012            Dr.T.V.Rao MD   1
BASIC TERMINOLOGY AND PRINCIPLES
• Symbiosis: Living together
• Commensalism: One symbiont
  benefits, other unaffected
• Mutualism: Both symbionts benefit
• Parasitism: One symbiont
 benefits, other is damaged
 12/10/2012     Dr.T.V.Rao MD    2
The Reality of Parasites
• 1.3 billion persons infected with
  Ascaris (1: 4 persons on earth)
• 300 million with
  Schistosomiasis
• 100 million new malaria cases/
  year
 12/10/2012    Dr.T.V.Rao MD     3
Laboratory Methods For Parasites In Faeces

• No technique is 100% successful in detecting
  parasites by a single stool examination, and at least
  three serial stools must be examined before a
  patient can be considered free from infections in
  which stages of parasites would be expected to be
  found in the faeces.
• Whilst clinical symptoms or a case history may
  provide clues as to which parasites may be
  present, each faecal specimen should be treated as
  an unknown, as parasite stages unrelated to the
  clinical picture may be present.
12/10/2012               Dr.T.V.Rao MD
Faecal specimens may contain several
          stages of Parasites
• Faecal specimens are examined for the presence of
  protozoa and helminthes larvae or eggs.
• The stages of protozoa found in stools are
  trophozoites and cysts. The stages of helminthes
  usually found in stools are eggs and larvae, though
  whole adult’s worms or segments of worms may also
  be seen. Adult worms and segments of tapeworms
  are usually visible to the naked eye, but
  eggs, larvae, trophozoites, and cysts can be seen only
  with the microscope.


12/10/2012               Dr.T.V.Rao MD
Collection of faecal specimens
1. Because of the fragile nature of many intestinal
   parasites, and the need to maintain their
   morphology for accurate identification, reliable
   microscopic diagnosis can’t be made unless the stool
   is collected properly.
2. Approximately 10 grams of fresh faeces
   uncontaminated by urine, oil, water, dyes or radio-
   opaque into a clean plastic container.
3. The container should be free from antiseptics and
   disinfectants.
4. Label all samples clearly with the patient’s
   name, reference number, date, and time of
   collection.
12/10/2012              Dr.T.V.Rao MD
Collection of faecal specimens
5. All samples should be accompanied by a
  requisition form from the physician
  giving relevant clinical details and recent
  travel history.
6. Samples and forms from patients with a
  confirmed or suspected diagnosis of
  certain infectious diseases such as AIDS
  or hepatitis should be clearly labeled
  with “Risk of Infection” or “Biohazard”

12/10/2012          Dr.T.V.Rao MD
Collection of faecal specimens
• 7.Most viable parasites are susceptible to
  desiccation or temperature variation. If time
  lapse between collection and observation is
  considerable, i.e. more than 4 days, it may be
  necessary to add some form of preservative to
  the faeces to retain the morphology as near to
  the original as possible.
• 8. Formed samples can be kept in a refrigerator at
  + 4c for a short while, but not in incubator.
• 9. Any whole worms or segments passed should
  be placed in a separate container

12/10/2012             Dr.T.V.Rao MD               8
Collect the Information of the
               Patient
• History (Age, occupation, residency, previous
  infection)
• Complaint                       Provisional diagnosis
• Clinical examination
• Investigations                  Confirm the diagnosis

    - Laboratory investigations
    - Radiology
    - Surgical intervention (Exploratory)

12/10/2012              Dr.T.V.Rao MD                 9
The Microscopy in Parasitology
• The Microscope is the parasitologist’s main
  tool. If possible the Microscope- should be
  binocular; most suitable objectives are the
  x10, x40, and x100.
• The Microscope must be covered and
  immersion oil removed from the lens -with
  xylene or ether when not in use.
• Calibration of the Microscope Eyepiece
  Micrometer:
   – On many occasions measuring the size of
     suspected parasites in faeces is helpful for
     identification.(eyepiece micrometer)
12/10/2012            Dr.T.V.Rao MD
Microscopic Examination of
              Wet Mount
• Wet mount is the simplest and easiest
  technique for the examination of faeces, and
  this method should be performed in all
  laboratories at the peripheral level.
• A wet mount can be prepared directly from
  faecal material or from concentrated
  specimens. The basic types of wet mount that
  should be used for each faecal examination
  are saline, iodine, and buffered methylene
  blue.

12/10/2012          Dr.T.V.Rao MD
The saline wet mount
• Is used for the initial microscopic
  examination of stools. It is employed
  primarily to demonstrate worm's
  eggs, larvae, protozoan trophozoites, and
  cysts.
• This type of mount can also reveal the
  presence of red blood cells and white
  blood cells.
12/10/2012         Dr.T.V.Rao MD
The Iodine wet mount
• Is used mainly to stain glycogen and the
  nuclei of cysts, if present. Cysts can
  usually be specifically identified in this
  mount.
• The buffered methylene blue (BMB) wet
  mount should be prepared each time
  amoebic trophozoites are seen in a saline
  wet mount, or when their presence is
  suspected.
12/10/2012         Dr.T.V.Rao MD
Direct saline and iodine mounts
1.   With a wax
     pencil writes
     the patient’s
     name or
     number and
     the date at the
     left-hand end
     of the slide.
12/10/2012         Dr.T.V.Rao MD
Preparing a faecal specimen
• Place a drop of saline in
  the centre of the left
  half of the slide and
  place a drop of iodine
  solution in the centre of
  the right half of the
  slide.
• Note: If the presence of
  amoebic trophozoites is
  suspected, warm saline
  (37c) should be used.


12/10/2012                   Dr.T.V.Rao MD   15
Preparation of Wet film
•    With an applicator
    stick (match or
    tooth pick), pick
    up a small portion
    of the specimen
    (size of a match
    head) and mix the
    drop of saline.
12/10/2012            Dr.T.V.Rao MD    16
STOOL EXAMINATION

                              MACROSCOPIC
                              EXAMINATION




     COLOUR        CONSISTENCY                   COMPOSITION     Adult PARASITES




                -Liquid (Troph)                                *Ascaris worm
                                          ?? Blood ?? Mucus
       Pale     -Formed (Cyst)                                 *E. vermicularis
                                          (dysentry)
                -Semi formed (Cyst)                            *T. saginata




12/10/2012                       Dr.T.V.Rao MD
DIAGNOSIS




        DIRECT      INDIRECT       MOLECULAR




         Urine
                      IHAT
         Stool
                       LAT
        Sputum                        PCR
                       IFAT
        Biopsy                     DNA probes
                      ELISA
         Blood
       Aspirates


12/10/2012         Dr.T.V.Rao MD
URINE
                                                EXAMINATION



                                       MACROSCOPIC      MICROSCOPIC



                      colour                               Sedimentation



             white             smoky                       concentration


                     Ether           Acetic acid
                                                              Membrane
               Chyluria           Blood
                                                               filtration
                Dissolve fat           RBC haemolysis

                                          S.
                     Filaria
12/10/2012
                      M.f         haematobium
                                    Dr.T.V.Raoova
                                    Clear MD
Examination
1. Put the slide with the mounts on the
  microscope stage and focus on the mount
  with the x10 or low-power objective.
2. Regulate the light in the microscope field with
  the sub stage diaphragm. You should be able
  to see objects in the field distinctly. Too much
  or too little light is not good.
3. Examine the entire coverslip area with the
  x10 objective; focus the objective on the top
  left-hand corner and move the slide
  systematically backwards and forwards, or up
  and down.
12/10/2012            Dr.T.V.Rao MD
Examination Cont.
4. When organisms or suspicious material are
  seen, switch to the high-dry objective, and
  increase the light by opening the sub stage
  diaphragm to observe the detailed
  morphology.
-This is a systematic examination. If mounts are
  examined in this way, any parasites present
  will usually be found. If the mount is not
  examined systematically, parasites may be
  missed. Examine each microscope field
  carefully, focusing up and down, before
  moving to the next field.

12/10/2012            Dr.T.V.Rao MD
12/10/2012   Dr.T.V.Rao MD
STOOL EXAMINATION
                    A Rapid Methods
         Saline smear                                        Iodine smear
                              saline                                           Iodine 1%




        Huge number of:
•Eggs                                                        Huge number of:

• Protozoal troph. Motility                            •Cyst morphological details

(Amoeb, flagellates)
 12/10/2012                            Dr.T.V.Rao MD                                 23
Need for Concentration Methods for
          Faecal examination
• A concentration
  procedure is performed
  mainly to separate the
  parasites from faecal
  debris. The concentration
  procedure not only
  increases the numbers of
  parasites in the sediment
  but it also unmasks
  them, making them more
  visible by removing
  organic and inorganic
  debris
12/10/2012               Dr.T.V.Rao MD   24
STOOL EXAMINATION
                      Scanty infection
                  Concentration techniques


     Sedimentation                                          Floatation

• Heavy eggs (Ascaris egg)                        • Non Operculated eggs

• Operculated eggs (Trematodes)                     Trematodes ( S. m.)

• Larvae (Strong sterc.)                            Cestode
                                                    Nematode(Hookworms,Trichostong)
• Cysts
                                                  • Cysts
    12/10/2012                    Dr.T.V.Rao MD                              25
STOOL EXAMINATION
                        Saline sedimentation



                                               Mesh wire gauze


    Saline   Emulsify
                                           Conical flask


10 g stool
                                        Sediment




12/10/2012                     Dr.T.V.Rao MD                     26
STOOL EXAMINATION
                          Kato technique
                                                Mesh screen




                                                                  Hole
                                                                  Template
 Remove the template



                                                              Cellophane soaked by glycerin
                                                                     (clears faeces




                                                               Egg count/ g stool




12/10/2012                      Dr.T.V.Rao MD                                           27
                            Egg quant. Of: Ascaris, T. trich., Hookworms, S. mansoni
Stool examination other
                    Texhniques
• Stoll’s technique for eggs of
  Ascaris,         T.
  trichiura., Hookworms, S. mansoni
• Baermann’s technique Detec. Of
  Nematode L. /stool, soil
• Cultures for Nematode larvae using Filter
  paper culture for Larvae of: St. stercoralis
  (A,L) and Hookworms
12/10/2012            Dr.T.V.Rao MD          28
Artifacts
• Artifacts other things, living or
  artificial, present in the stool that are
  not parasites and could mislead the
  laboratory worker.
• Note: “Artifacts not to be mistaken
  for cysts”.

12/10/2012         Dr.T.V.Rao MD
12/10/2012   Dr.T.V.Rao MD
Plant cells                                                                       Pollen grains



Air bubble
                                                Plant    fibre
                                                                                Plant hairs




                                                     Non-human
                                                coccidial oocysts                                       Fat droplets




Soapy plaques
                                  Starch cell                    Charcot leyden crystals                   Muscle fibers




          Fatty acids                             Macrophage
       12/10/2012                                       Dr.T.V.Rao MD              Epithelial cells
Examining other Specimens




12/10/2012       Dr.T.V.Rao MD     32
SPUTUM
                    EXAMINATION



        MACROSCOPIC             MICROSCOPIC
                                                                       NaOH

             Appearance         Concentration                          Sputum


                                                                Centrifuge
               Bloody (Parag)



               Rusty brown
                 (Parag)




12/10/2012                                      Dr.T.V.Rao MD
BIOPSY
                    SPECIMEN


                    MUSCLE              RECTAL
             SKIN
                    BIOPSY              BIOPSY


    O. Volvulus     T. Spiralis       Schistosomes
        mf            larvae               egg


12/10/2012            Dr.T.V.Rao MD
•Lumbar puncutre
• Centrifuge
•Examine sed.




 12/10/2012        Dr.T.V.Rao MD
BLOOD
                                          EXAMINATION


                                   Buffy coat                            Knott’s conc.
             Blood films                                 QBC technique
                                     films                                  tech.



 Thin                      Thick




12/10/2012                               Dr.T.V.Rao MD
BLOOD EXAMINATION
                   BLOOD FILMS
             • Thin                                       • Thick
 Bld drop

                      spread                                          Circular motion




                       Air dry                                         Air dry




                      methyl alcohol
                                                                       Geimsa

                        Geimsa
                                                 Malaria, Babesia, Filaria, Tryp.
12/10/2012                       Dr.T.V.Rao MD                                   37
Observe the Thin and Thick Smear




12/10/2012    Dr.T.V.Rao MD    38
QBC Method is used in ….
• The QBC Malaria method is the simplest and
  most sensitive method for diagnosing the
  following diseases.
• Malaria
• Babesiosis
• Trypanosomiasis (Chagas disease, Sleeping
  Sickness)
• Filariasis (Elephantiasis, Loa-Loa)
• Relapsing Fever (Borreliosis)

12/10/2012           Dr.T.V.Rao MD             39
QBC A QUICKER ALTERNATIVE IN
                  MALARIA




12/10/2012          Dr.T.V.Rao MD      40
How to read the QBC results ….
• When the operator looks
  through the wall of the
  tube, the nucleus of the
  parasite fluoresces bright
  green, and the cytoplasm
  shows up as yellow-orange.
  The shape and colours are
  quite characteristic, and
  since the parasites are
  concentrated up to
  1000X, there are usually a
  large number of them in
  any field of view in this area
  of the tube.


12/10/2012                    Dr.T.V.Rao MD   41
URINE EXAMINATION
        SEDIMENTATION                              CONCENTRATION
                 Clean conical
                glass receptacle




                15-20 min                                  Centrifuge (2 min)




12/10/2012                         Dr.T.V.Rao MD                       42
URINE EXAMINATION
             Membrane filtration technique

                     air


               10 ml urine


       Nucleopore filter



                             + Saline




                                                 Eggs of Schistosoma
12/10/2012                       Dr.T.V.Rao MD                         43
URINE EXAMINATION




          HELMINTHES       PROTOZOA               ARTHROPODES




•   S. haem.egg
•   E. vermic. egg
                                            • Pthirus pubis
•   S. mansoni egg        • T. Vag troph
•   Mf (Ov, Wb)
•   H sand




    12/10/2012              Dr.T.V.Rao MD
Indirect immunological diagnosis
• Serology – All tests
  available
                                      • Skin Tests –
      – IHA                             Specificity low,
      – ELISA
• More useful in
                                        cross reactions
      –      Amoebiasis                 common
      –      Leishmaniasis
      –      Malaria
                                              –Casoni’s test
      –      Toxoplasmosis                    –Leishmanin test
      –      Trichinosis
      –      Filariasis
      –      Echinococcosis
12/10/2012                    Dr.T.V.Rao MD                      45
• Programme Created by Dr.T.V.Rao MD
   for Basic learning in Human Parasitology
     for Undergraduate Medical Students
                     • Email
             • doctortvrao@gmail.com


12/10/2012            Dr.T.V.Rao MD       46

More Related Content

PARASITIC INFECTIONS

  • 1. PARASITIC INFECTIONS BASICS IN DIAGNOSIS Dr.T.V.Rao MD 12/10/2012 Dr.T.V.Rao MD 1
  • 2. BASIC TERMINOLOGY AND PRINCIPLES • Symbiosis: Living together • Commensalism: One symbiont benefits, other unaffected • Mutualism: Both symbionts benefit • Parasitism: One symbiont benefits, other is damaged 12/10/2012 Dr.T.V.Rao MD 2
  • 3. The Reality of Parasites • 1.3 billion persons infected with Ascaris (1: 4 persons on earth) • 300 million with Schistosomiasis • 100 million new malaria cases/ year 12/10/2012 Dr.T.V.Rao MD 3
  • 4. Laboratory Methods For Parasites In Faeces • No technique is 100% successful in detecting parasites by a single stool examination, and at least three serial stools must be examined before a patient can be considered free from infections in which stages of parasites would be expected to be found in the faeces. • Whilst clinical symptoms or a case history may provide clues as to which parasites may be present, each faecal specimen should be treated as an unknown, as parasite stages unrelated to the clinical picture may be present. 12/10/2012 Dr.T.V.Rao MD
  • 5. Faecal specimens may contain several stages of Parasites • Faecal specimens are examined for the presence of protozoa and helminthes larvae or eggs. • The stages of protozoa found in stools are trophozoites and cysts. The stages of helminthes usually found in stools are eggs and larvae, though whole adult’s worms or segments of worms may also be seen. Adult worms and segments of tapeworms are usually visible to the naked eye, but eggs, larvae, trophozoites, and cysts can be seen only with the microscope. 12/10/2012 Dr.T.V.Rao MD
  • 6. Collection of faecal specimens 1. Because of the fragile nature of many intestinal parasites, and the need to maintain their morphology for accurate identification, reliable microscopic diagnosis can’t be made unless the stool is collected properly. 2. Approximately 10 grams of fresh faeces uncontaminated by urine, oil, water, dyes or radio- opaque into a clean plastic container. 3. The container should be free from antiseptics and disinfectants. 4. Label all samples clearly with the patient’s name, reference number, date, and time of collection. 12/10/2012 Dr.T.V.Rao MD
  • 7. Collection of faecal specimens 5. All samples should be accompanied by a requisition form from the physician giving relevant clinical details and recent travel history. 6. Samples and forms from patients with a confirmed or suspected diagnosis of certain infectious diseases such as AIDS or hepatitis should be clearly labeled with “Risk of Infection” or “Biohazard” 12/10/2012 Dr.T.V.Rao MD
  • 8. Collection of faecal specimens • 7.Most viable parasites are susceptible to desiccation or temperature variation. If time lapse between collection and observation is considerable, i.e. more than 4 days, it may be necessary to add some form of preservative to the faeces to retain the morphology as near to the original as possible. • 8. Formed samples can be kept in a refrigerator at + 4c for a short while, but not in incubator. • 9. Any whole worms or segments passed should be placed in a separate container 12/10/2012 Dr.T.V.Rao MD 8
  • 9. Collect the Information of the Patient • History (Age, occupation, residency, previous infection) • Complaint Provisional diagnosis • Clinical examination • Investigations Confirm the diagnosis - Laboratory investigations - Radiology - Surgical intervention (Exploratory) 12/10/2012 Dr.T.V.Rao MD 9
  • 10. The Microscopy in Parasitology • The Microscope is the parasitologist’s main tool. If possible the Microscope- should be binocular; most suitable objectives are the x10, x40, and x100. • The Microscope must be covered and immersion oil removed from the lens -with xylene or ether when not in use. • Calibration of the Microscope Eyepiece Micrometer: – On many occasions measuring the size of suspected parasites in faeces is helpful for identification.(eyepiece micrometer) 12/10/2012 Dr.T.V.Rao MD
  • 11. Microscopic Examination of Wet Mount • Wet mount is the simplest and easiest technique for the examination of faeces, and this method should be performed in all laboratories at the peripheral level. • A wet mount can be prepared directly from faecal material or from concentrated specimens. The basic types of wet mount that should be used for each faecal examination are saline, iodine, and buffered methylene blue. 12/10/2012 Dr.T.V.Rao MD
  • 12. The saline wet mount • Is used for the initial microscopic examination of stools. It is employed primarily to demonstrate worm's eggs, larvae, protozoan trophozoites, and cysts. • This type of mount can also reveal the presence of red blood cells and white blood cells. 12/10/2012 Dr.T.V.Rao MD
  • 13. The Iodine wet mount • Is used mainly to stain glycogen and the nuclei of cysts, if present. Cysts can usually be specifically identified in this mount. • The buffered methylene blue (BMB) wet mount should be prepared each time amoebic trophozoites are seen in a saline wet mount, or when their presence is suspected. 12/10/2012 Dr.T.V.Rao MD
  • 14. Direct saline and iodine mounts 1. With a wax pencil writes the patient’s name or number and the date at the left-hand end of the slide. 12/10/2012 Dr.T.V.Rao MD
  • 15. Preparing a faecal specimen • Place a drop of saline in the centre of the left half of the slide and place a drop of iodine solution in the centre of the right half of the slide. • Note: If the presence of amoebic trophozoites is suspected, warm saline (37c) should be used. 12/10/2012 Dr.T.V.Rao MD 15
  • 16. Preparation of Wet film • With an applicator stick (match or tooth pick), pick up a small portion of the specimen (size of a match head) and mix the drop of saline. 12/10/2012 Dr.T.V.Rao MD 16
  • 17. STOOL EXAMINATION MACROSCOPIC EXAMINATION COLOUR CONSISTENCY COMPOSITION Adult PARASITES -Liquid (Troph) *Ascaris worm ?? Blood ?? Mucus Pale -Formed (Cyst) *E. vermicularis (dysentry) -Semi formed (Cyst) *T. saginata 12/10/2012 Dr.T.V.Rao MD
  • 18. DIAGNOSIS DIRECT INDIRECT MOLECULAR Urine IHAT Stool LAT Sputum PCR IFAT Biopsy DNA probes ELISA Blood Aspirates 12/10/2012 Dr.T.V.Rao MD
  • 19. URINE EXAMINATION MACROSCOPIC MICROSCOPIC colour Sedimentation white smoky concentration Ether Acetic acid Membrane Chyluria Blood filtration Dissolve fat RBC haemolysis S. Filaria 12/10/2012 M.f haematobium Dr.T.V.Raoova Clear MD
  • 20. Examination 1. Put the slide with the mounts on the microscope stage and focus on the mount with the x10 or low-power objective. 2. Regulate the light in the microscope field with the sub stage diaphragm. You should be able to see objects in the field distinctly. Too much or too little light is not good. 3. Examine the entire coverslip area with the x10 objective; focus the objective on the top left-hand corner and move the slide systematically backwards and forwards, or up and down. 12/10/2012 Dr.T.V.Rao MD
  • 21. Examination Cont. 4. When organisms or suspicious material are seen, switch to the high-dry objective, and increase the light by opening the sub stage diaphragm to observe the detailed morphology. -This is a systematic examination. If mounts are examined in this way, any parasites present will usually be found. If the mount is not examined systematically, parasites may be missed. Examine each microscope field carefully, focusing up and down, before moving to the next field. 12/10/2012 Dr.T.V.Rao MD
  • 22. 12/10/2012 Dr.T.V.Rao MD
  • 23. STOOL EXAMINATION A Rapid Methods Saline smear Iodine smear saline Iodine 1% Huge number of: •Eggs Huge number of: • Protozoal troph. Motility •Cyst morphological details (Amoeb, flagellates) 12/10/2012 Dr.T.V.Rao MD 23
  • 24. Need for Concentration Methods for Faecal examination • A concentration procedure is performed mainly to separate the parasites from faecal debris. The concentration procedure not only increases the numbers of parasites in the sediment but it also unmasks them, making them more visible by removing organic and inorganic debris 12/10/2012 Dr.T.V.Rao MD 24
  • 25. STOOL EXAMINATION Scanty infection Concentration techniques Sedimentation Floatation • Heavy eggs (Ascaris egg) • Non Operculated eggs • Operculated eggs (Trematodes) Trematodes ( S. m.) • Larvae (Strong sterc.) Cestode Nematode(Hookworms,Trichostong) • Cysts • Cysts 12/10/2012 Dr.T.V.Rao MD 25
  • 26. STOOL EXAMINATION Saline sedimentation Mesh wire gauze Saline Emulsify Conical flask 10 g stool Sediment 12/10/2012 Dr.T.V.Rao MD 26
  • 27. STOOL EXAMINATION Kato technique Mesh screen Hole Template Remove the template Cellophane soaked by glycerin (clears faeces Egg count/ g stool 12/10/2012 Dr.T.V.Rao MD 27 Egg quant. Of: Ascaris, T. trich., Hookworms, S. mansoni
  • 28. Stool examination other Texhniques • Stoll’s technique for eggs of Ascaris, T. trichiura., Hookworms, S. mansoni • Baermann’s technique Detec. Of Nematode L. /stool, soil • Cultures for Nematode larvae using Filter paper culture for Larvae of: St. stercoralis (A,L) and Hookworms 12/10/2012 Dr.T.V.Rao MD 28
  • 29. Artifacts • Artifacts other things, living or artificial, present in the stool that are not parasites and could mislead the laboratory worker. • Note: “Artifacts not to be mistaken for cysts”. 12/10/2012 Dr.T.V.Rao MD
  • 30. 12/10/2012 Dr.T.V.Rao MD
  • 31. Plant cells Pollen grains Air bubble Plant fibre Plant hairs Non-human coccidial oocysts Fat droplets Soapy plaques Starch cell Charcot leyden crystals Muscle fibers Fatty acids Macrophage 12/10/2012 Dr.T.V.Rao MD Epithelial cells
  • 33. SPUTUM EXAMINATION MACROSCOPIC MICROSCOPIC NaOH Appearance Concentration Sputum Centrifuge Bloody (Parag) Rusty brown (Parag) 12/10/2012 Dr.T.V.Rao MD
  • 34. BIOPSY SPECIMEN MUSCLE RECTAL SKIN BIOPSY BIOPSY O. Volvulus T. Spiralis Schistosomes mf larvae egg 12/10/2012 Dr.T.V.Rao MD
  • 35. •Lumbar puncutre • Centrifuge •Examine sed. 12/10/2012 Dr.T.V.Rao MD
  • 36. BLOOD EXAMINATION Buffy coat Knott’s conc. Blood films QBC technique films tech. Thin Thick 12/10/2012 Dr.T.V.Rao MD
  • 37. BLOOD EXAMINATION BLOOD FILMS • Thin • Thick Bld drop spread Circular motion Air dry Air dry methyl alcohol Geimsa Geimsa Malaria, Babesia, Filaria, Tryp. 12/10/2012 Dr.T.V.Rao MD 37
  • 38. Observe the Thin and Thick Smear 12/10/2012 Dr.T.V.Rao MD 38
  • 39. QBC Method is used in …. • The QBC Malaria method is the simplest and most sensitive method for diagnosing the following diseases. • Malaria • Babesiosis • Trypanosomiasis (Chagas disease, Sleeping Sickness) • Filariasis (Elephantiasis, Loa-Loa) • Relapsing Fever (Borreliosis) 12/10/2012 Dr.T.V.Rao MD 39
  • 40. QBC A QUICKER ALTERNATIVE IN MALARIA 12/10/2012 Dr.T.V.Rao MD 40
  • 41. How to read the QBC results …. • When the operator looks through the wall of the tube, the nucleus of the parasite fluoresces bright green, and the cytoplasm shows up as yellow-orange. The shape and colours are quite characteristic, and since the parasites are concentrated up to 1000X, there are usually a large number of them in any field of view in this area of the tube. 12/10/2012 Dr.T.V.Rao MD 41
  • 42. URINE EXAMINATION SEDIMENTATION CONCENTRATION Clean conical glass receptacle 15-20 min Centrifuge (2 min) 12/10/2012 Dr.T.V.Rao MD 42
  • 43. URINE EXAMINATION Membrane filtration technique air 10 ml urine Nucleopore filter + Saline Eggs of Schistosoma 12/10/2012 Dr.T.V.Rao MD 43
  • 44. URINE EXAMINATION HELMINTHES PROTOZOA ARTHROPODES • S. haem.egg • E. vermic. egg • Pthirus pubis • S. mansoni egg • T. Vag troph • Mf (Ov, Wb) • H sand 12/10/2012 Dr.T.V.Rao MD
  • 45. Indirect immunological diagnosis • Serology – All tests available • Skin Tests – – IHA Specificity low, – ELISA • More useful in cross reactions – Amoebiasis common – Leishmaniasis – Malaria –Casoni’s test – Toxoplasmosis –Leishmanin test – Trichinosis – Filariasis – Echinococcosis 12/10/2012 Dr.T.V.Rao MD 45
  • 46. • Programme Created by Dr.T.V.Rao MD for Basic learning in Human Parasitology for Undergraduate Medical Students • Email • doctortvrao@gmail.com 12/10/2012 Dr.T.V.Rao MD 46