Laboratory Diagnostics

Question 1

Uroliths 01

The veterinary surgeon has recently obtained a urine sample from a patient with signs of cystitis. The sample has been centrifuged and you are requested to prepare this sample for examination under the microscope to examine the sediment.
A. Make and prepare the slide for examination
B. Using this microscope provided, find a crystal and place it in the centre of the field of view (please note: you are not expected to identify this crystal)
C. Identify the crystal shown in the photograph

Answer 1

1. Select a pipette
2. Select a microscope slide
3. Wear gloves
4. Remove most of the supernatant fluid and dispose of correctly
5. Sediment not disturbed
6. Re-suspend the sediment in remaining supernatant fluid (by flicking the tube or gently shaking it)
7. Pipette up a small amount of remaining supernatant and sediment
8. Place sufficient sample onto microscope slide
9. Carefully place a cover slip over sample minimising the risk of air bubbles
10. Label slide
11. Dispose of used pipette, urine and used materials into clinical waste bin
12. Slide prepared correctly for examination (e.g. supernatant removed, crystals not damaged)
13. Safe practice: gloves worn and no contamination of self with urine
14. Place microscope slide on stage, correct way up
15. Look at the stage directly whilst racking it up so that it is positioned just below the objective lens
16. Look don the eyepieces
17. Adjust the coarse and fine focus to focus the microscope
18. Methodically scan the area of slide
19. Locate a crystal
20. Correctly identify urine crystals in the photograph

Question 2

PCV 02
You are nursing a patient with anaemia
The veterinary surgeon requests that you obtain a daily PCV for this patient
A. Prepare the blood sample and set up the centrifuge for this test
B. Obtain the PCV reading using the Hawksley haematocrit reader from the pre-prepared sample provided. Record your results including units.

Note: you are not expected to start the centrifuge

Answer 2

1. Wear gloves
2. Select EDTA sample
3. Mix sample gently
4. Remove 1 or 2 plain capillary tubes
5. Insert capillary tube into sample (holding the sample tube at an angle)
6. Fill the tube to at least 3/4 full
7. Place finger over top end of tube or keep tube horizontal
8. Remove from sample
9. Wipe outside of capillary tube with a tissue
10. Plug end of capillary tube with soft clay sealant
11. Place capillary tube into mictohaematocrit centrifuge
12. Clay plug is against outer rim
13. Balance centrifuge
14. Screw inner safety lid down over samples
15. Close and lock main lid
16. Set at 10,000 rpm (or fast setting, depending on make of centrifuge)
17. For 5 minutes
18. Dispose of any used capillary tubes into glass waste (if applicable) and other used materials into clinical waste bin
19. Capillary tube prepared to produce PCV of diagnostic quality
20. Safe practice: gloved worn and no contamination of self or equipment with blood
21. PCV reading: place prepared capillary tube into PCV reader
22. Sealed end at the bottom
23. Bottom of red blood cell (RBC) layer is at the zero line or the reader
24. Top of plasma is at the 100% line of the reader
25. Move the adjustable PCV reading line to intersect the top of the RBC layer
26. Record PCV results correctly as a percentage

Question 3

What setting should the centrifuge be at for a PCV sample?

Answer 3

10,000 RPM for 5 minutes (or fast setting)

Question 4

Where should the bottom of the RBC layer be placed on the Hawksley reader for a PCV?

Answer 4

At the zero line

Question 5

Where should the top of the plasma layer the positioned on the Hawksley reader for a PCV?

Answer 5

At the 100% line

Question 6

Urinalysis 03
You are nursing a patient with dysuria. Using the equipment available and the urine sample provided, obtain and record the following parameters:
A. A refractometer urine specific gravity reading
B. A dipstick reading

Answer 6

1. Place 2-3 drops of distilled water on the prism surface of the refractometer
2. Hold the refractometer up to light source and look down eye piece
3. Calibrate the refractometer to 1.000 on the USG (or W) scale
4. Lift cover and dry prism surface using a dry tissue
5. Wear gloves
6. Invert the tube to gently mix the urine sample
7. Pipette 1-2 drops of urine onto the prism surface
8. Close the cover
9. Hold up refractometer and look down eyepiece
10. Read and record the actual urine specific gravity reading, having correctly calibrated refractometer
11. Rinse the prism with water
12. Dry the prism
13. Select urine dipstick test strips
14. Remove one test strip
15. Replace lid immediately
16. Cover test strip pads with urine
17. Immediately note the time
18. Wait for appropriate length of time
19. Read and record dipstick measurements correctly
20. Dispose of used dipstick and used materials into clinical waste
21. Safe practice: gloves worm and no contamination of self with urine

Question 7

Blood smear 04

You are nursing a patient that is showing signs of anaemia.
The veterinary surgeon has asked you to prepare and stain a blood smear from a blood sample which has just been obtained from the patient.
A. Demonstrate how you would prepare the smear
B. Check and comment on the quality of the smear you have prepared
C. Stain the blood smear provided by the examiner using the Diff-Quick solutions

Answer 7

1. Wear gloves
2. Select microscope slide
3. Select spreader slide
4. Select EDTA sample
5. Gently mix sample
6. Select plain capillary tube
7. Insert tube into blood sample
8. Draw up small amount of blood into the tube
9. Place a finger over top of tube or keep tube horizontal to prevent leakage of blood
10. Remove tube from sample
11. Dot a small amount of blood near one end of slide
12. Discard capillary tube into clinical glass waste bin
13. Clean and dry the spreader surface of the slide
14. Hold blood sample slide firmly on the work surface
15. Place spreader on opposite end of the slide to the blood
16. Draw spreader back to the drop of blood
17. Allow blood to spread along edge of spreader
18. Push spreader away from blood drop
19. Use single, smooth motion
20. Smear produced of reasonable diagnostic quality (EDTA sample selected and smear with edge and tail)
21. Rapidly air dry the slide
22. Label the slide
23. Check and comment on the quality of the smear
24. Diff-quick technique: slide dipped into the fixative (pale blue)
25. Dipped 5 times for one second each time
26. Excess fluid allowed to drip back into the jar
27. Slide dipped into stain solution 1 (red)
28. Dipped 5 times for one second each time
29. Excess fluid allowed to drip back into the jar
30. Slide dipped into stain solution 2 (purple)
31. Dipped 5 times for one second each time
32. Excess fluid allowed to drip back into the jar
33. Slide ringed with distilled water
34. Slight placed vertically and left to dry
35. Smear correctly stained
36. Safe practice: gloved worn with no contamination of self with blood sample or stain solutions

Question 8

Parasite 05
The veterinary surgeon is consulting and has passed you a microscope slide. The vet has asked you to examine it under the microscope for any evidence of parasites.
A. Set up the microscope, locate and focus on the parasite on the prepared slide
B. Write down the vernier scale reading to relocate the parasite is the slide is moved
C. Identify the ectoparasites on the photographs provided by the examiner

Answer 8

1. Check that the light intensity switch (rheostat) is turned to the lowest setting to avoid damaging the build
2. Turn on the microscope
3. Rack down the mechanical stage to as low as possible
4. Select the lowest power objective lense(x4 or x10 depending on the microscope available)
5. Place microscope slide on stage, correct way up
6. Look at the stage directly whilst racking it up so that’s it is positioned just below the objective lens (or rack down eyepiece to the stage, depending on the microscope)
7. Look down the eyepieces
8. Adjust the rheostat to a medium setting
9. Adjust the height of the sub stage condenser to a few millimetres below the stage
10. Adjust the fine/coarse focus in order to focus the microscope
11. Focus on the parasite on the prepared slide
12. Read and correctly record the horizontal scale
13. Read and correctly record the vertical scale
14. Identify parasite A (common name/latin name)
15. Identify parasite B (common name/Latin name)

Question 9

Blood sample 06
The veterinary surgeon has requested that you obtain a blood sample via the jugular even for routine haematology from a feline patient.
A. Demonstrate hand hygiene using the world health organisation (WHO) 6 step method and the alcohol gel provided
B. Select and prepare the equipment required for obtaining the blood sample
Place the equipment in the tray, as indicated by the examiner

Answer 9

1. Remove all hand and wrist jewellery, roll sleeve up (candidates without jewellery or long sleeves awarded this step)
2. Nails well manicured, cleaned and unvarnished. False nails are not permitted
3. Apply 3-5 mls alcohol rub to palms (usually 1 pump)
4. Rub hands palm to palm (minimum 3 strokes)
5. Rub right palm over the back of the left hand with interlaced fingers (minimum 3 strokes)
6. Rub left palm over back of right hand with interlaces fingers (minimum 3 strokes)
7. Palm to palm with interlaced fingers (minimum 3 strokes)
8. Backs of fingers to opposing palms with finer interlocked (minimum 3 strokes)
9. Rotationally rub the left thumb clasped in the right palm (minimum 3 strokes)
10. rotationally rub the right thumb clasped in the left palm (minimum 3 strokes)
11. Rotationally rub finger tips of right hand in left palm (minimum of 3 strokes)
12. Rotationally rub finger tips of left hand in right palm (minimum 3 strokes)
13. Air dry hands or secure hands are dry before selecting equiptment
14. Effective hand hygiene technique used (WHO method)
15. Select a pair of gloves
16. Select clippers
17. Select clipper blades
18. Apply clipper blades correctly
19. Check clippers are functioning
20. Select appropriate antiseptic skin preparation swab
21. Select swab for post venipuncture
22. Select appropriate needle
23. Select appropriate syringe
24. Assemble syringe and needle
25. Correct syringe and needle assembled aseptically, without risk of needle stick injury
26. Pre loose plunger of syringe
27. Select appropriate blood sample tube
28. Label sample tube appropriately (patient name and date)

Question 10

Ear Swab 07

The veterinary surgeon has requested that an aural swab is taken for bacterial culture by an external laboratory.

Using the equipment provided, take and aseptic sample from the x ear of this patient.

Package the sample to meet UN packaging instructions P650, ready for dispatch to the external laboratory.
This will be collected by courier.
Please note: you have already washed your hands

Answer 10

1. Select charcoal swab
2. Ask for assistance and give instruction for the patient to be restrained
3. Select and wear gloved prior to commencing the procedure
4. Using the swab, collect sample from the ear without contamination
5. Collect sufficient sample for analysis by gently rotating swab to cover all surfaces
6. Patient handled and sample collected without causing trauma to the ear
7. Replace swab into the cover tube and secure
8. Remove gloved and dispose of correctly
9. Label swab with the location sample is being collected from (e.g. left ear)
10. Label swab with the animal’s name/ID, owners name and the date
11. Sample correctly collected and labelled using a safe, effective and aseptic technique
12. Insert client’s name on laboratory form
13. Insert animals details on laboratory form
14. Sample type submitted on laboratory form
15. Insert test required on laboratory form
16. Wrap swab in absorbent material
17. Wrap swab in bubble wrap
18. Place swab in polythene bag
19. Remove air from polythene bag and seal
20. Place laboratory form in polythene bag (separated from sample)
21. Place form and sample into outer packaging
22. Secure outer packaging
23. Place completed packaging in courier tray - ready for submission
24. Sample packed to meet United Nations packaging instructions P650
25. Contaminated gloves not worn during completion of steps 8-23

Question 11

Wound Swab 08
The veterinary surgeon has requested that a swab is taken for bacterial culture by an external laboratory.
Using the equipment provided, take an aseptic sample from the X carpus of this patient. Package the sample to meet UN packaging instruction P650, ready for dispatch to the external laboratory.
This will be collected by courier.
Please note: you have already washed your hands

Answer 11

1. Select charcoal swab
2. Select and wear gloved prior to commencing the procedure
3. Using the swab, collect sample rom the wound without contamination
4. Collect sufficient sample for analysis by gently rotating swab to cover all surfaced
5. Replace swab into the cover tube and secure
6. Sample collected without causing further trauma to the patient
7. Remove gloved and dispose of correctly
8. Label swab with the location sample is being collected from (e.g. right carpus)
9. Label swab with the animal’s name/ID, owner’s name and the date
10. Sample correctly collected and labelled using a safe, effective and aseptic techniques
11. Insert client’s name on laboratory form
12. Insert animals details on laboratory form.
13. Sample type submitted on laboratory form
14. Identify test required on laboratory form
15. Wrap swab in absorbent material
16. Wrap swab in bubble wrap
17. Place swab in polythene bag
18. Remove air from polythene bag and seal
19. Place laboratory form in polythene bag (separated from sample)
20. Place form and sample into outer packaging
21. Secure outer packaging
22. Place completed package in courier tray - ready for submission
23. Sample packed to meet United Nations packaging instructions P650
24. Contaminated gloved not worn using completion of steps 8-22