This document summarizes an in-service training on urinalysis or urine cytobacteriological tests (ECBU) presented by Nkengacha Marcellous Agendia. The training covered the meaning and aim of ECBU, urine collection methods, microscopic examination including identification of cells, crystals and casts, biochemical testing, gram staining and bacterial culture. The goal was to diagnose urinary tract infections and properly interpret normal versus abnormal ECBU results.

1. In-Service Training
Laboratory Section, District Hopital
Sangmelima
Date: January 23rd, 2013
Venue: Laboratory hall
Time: 2:30 pm
Coordinator/presenter: Nkengacha Marcellous Agendia

2. In-Service Training
E C B UBy
Nkengacha Marcellous Agendia
Biomedical Engineer

3. In-Service Training
1. Meaning & Aim of test
2. The Urinary System
3. Collection methods
4. Procedure
5. Results
6. Interpretation (normal/abnormal)
ECBU

4. In-Service Training
 Examen Cyto-Bacteriologie
d’Urine
 Urine Cytobacteriological Test
or Urinalysis in English
ECBU

5. In-Service Training
Cyto (-logy)  cells study
Bacteriologie  bacterial study
ECBU

6. In-Service Training
1. Diagnose UTI
 Urethral
 Bladder
 Ureter
 Nephrones
 Glomerulose
ECBU

7. In-Service Training
1. Diagnose UTI
 Urethral
 Bladder
 Ureter
 Nephrones
 Glomerulose
ECBU

8. In-Service Training
2. Treatment followup
3. Other kidney diseases
ECBU

9. In-Service Training
ECBU

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 Catheterized
 Suprapubic urine
 Clean catch midstream
 Adapted urine kits for children
ECBU

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Catheterized
 Patients with catheter
 Risk of contamination high
ECBU

12. In-Service Training
Suprapubic urine
 Contamination risk is zero.
 Involves passage of a needle via the
pubic region.
 It is the best option as far as urine
collection is concerned
 Induces pain
ECBU

13. In-Service Training
Adapted urine kits for children
 Designed for urine collection in
children.
 There is some risk of contamination
 It is more convenient
ECBU

14. In-Service Training
Clean catch midstream
 Most widely used method
 Convenient for the patient
 Risk of contamination from skin,
vagina and normal floral
 The middle portion of urine often
required
ECBU

15. In-Service Training
 Random
 First urine of the day
ECBU

16. In-Service Training
 Vagina
 Skin
 Urethral normal floral
 Environment
ECBU

17. In-Service Training
Urethral normal floral are mainly;
 Staphylococcus
 Micrococcus
 Enterobacterea
 Corynebacteria
 Non-groupable Streptococcus
ECBU

18. In-Service Training
Urethral normal floral
 These are both Gram(+) and Gram(-)
 Often found in first portion of urine
and sperm
 This can be avoided by cleaning and
collection of second portion
ECBU

19. In-Service Training
 Labelled
 Clean
 Sterile
 Dry
 Transparent
 Wide-mouth container
ECBU

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 Examine immediately or
 Note macroscopy and preserve in a
refrigerator
 T°C = 2 – 8°C
ECBU

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 Colour
 Transparency
 Others (formed physical elements)
ECBU

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Colour;
 Colourless
 Yellow (dark yellow)
 Coca-cola
 Red, etc…
ECBU

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Transparency;
(Ability to read a text via the sample);
 Clear
 Cloudy (lightly cloudy)
ECBU

24. In-Service Training
ECBU

25. In-Service Training
Biochemical tests with Combi 10 include;
 Nitrite
 Leukocyte esterase
 Blood
 Protein
ECBU

26. In-Service Training
Nitrite Vs Leukocyte esterase:
 Negative tests for both exclude UTI
 ECBU may not be useful
 Positive for one or both indicate UTI
 ECBU &/or culture is strongly
recommended
ECBU

27. In-Service Training
Blood:
 Suspect S. hematobium in an endemic
area
 Especially for children between 5 – 15
years
 A concomittant UTI is possible
ECBU

28. In-Service Training
ECBU

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 Done after macroscopy
 Done to harvest cells and other elements
 Usually for about 1000r/5mins.
ECBU

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To identify elements and rate their quantity;
 Cells
 Crystal
 Casts
 Parasites
ECBU

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Cells;
 WBCs;
ECBU

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Cells;
 RBCs
ECBU

33. In-Service Training
Cells;
 Yeast cells
ECBU

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Cells;
 Epithelial cells
1. Transitional EC
Found in;
Ureter, bladder,
part of urethral
ECBU

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Cells;
 Epithelial cells
2. Squamous EC
Found on the;
Skin, and vagina.
contaminant
ECBU

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Crystals;
 Calcium oxalate
 Calcium carbonate
 Amorphous
 Ammonium biurate
 Bilirubin
 Magnesium ammonium phosphate
ECBU

37. In-Service Training
Cystals;
 Calcium oxalate
ECBU

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Cystals;
 Amorphous
ECBU

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Cystals;
 Magnesium
ammonium
phosphate
ECBU

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Parasites;
 Schistosoma hematobium
 Pinworm (very rare)
ECBU

41. In-Service Training
 Make a wet mount
 Overlay with a cover slip
 Put on a microscopic stage
 Focus with objective 10X
 Identify elements with 40X
ECBU

42. In-Service Training
Using objective 10X (LPF);
 Good in doing focus
 Good for searching elements
 Covers more field of view than 40X
 Magnifies elements up to 10X normal size
 Most elements are not very clear
 Always switch to 40X for better view
ECBU

43. In-Service Training
Using objective 40X (HPF);
 Not very good for focus
 Good for identification
 Covers smaller field of view than 10X
 Magnifies elements up to 40X normal size
 Most elements are very clear
 End examination with this objactive
ECBU

44. In-Service Training
Leukocytes and RBCs;
 Average of total number of cells counted
in ‘X’ fields, usually 10 fields.
 Report number of cells per HPF (40X).
 Results with decimal create a range
ECBU

45. In-Service Training
Leukocytes and RBCs - Example;
Field 1 = 4 RBCs
Field 2 = 4 RBCs
Field 3 = 5 RBCs
Result = (4+4+5)/3
= 4.3 cells
Actual result = 4 – 5 cells/HPF
ECBU

46. In-Service Training
Casts;
 The same as leukocytes and RBCs
 But reported per LPF (10X) and not HPF.
ECBU

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Others;
 Rare,
 Few,
 Moderate or
 Many per HPF.
ECBU

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ECBU

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 It is not a routine step
 Done when Combi 10 and/or
wetmount are abnormal.
 To identify bacteria type;
 Gram (+) or Gram (-)
 Read with 100X objective
ECBU

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 Gram Positive bacteria  Purple colour
 Gram Negative bacteria  Pink colour
ECBU

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 Plus signs (+, ++)
 Rare, few, moderate, many
ECBU

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 To identify the species of bacteria.
 To select an appropriate antibiotic.
 Estimates the concentration of bacteria
 Not done routinely.
ECBU

53. In-Service Training
Biochemistry;
 Nitrite  Negative
 Leukocyte esterase  Negative(0 – 5/HPF)
Wetmount;
 Leukocytes  0 – 5/HPF
 RBCs  0 – 2/HPF
 Parasites  Negative
ECBU

54. In-Service Training
Wetmount;
 Bacteria  depends on;
 Collection method
 Level of leukocytes present
 Epithelial cells  depends on type;
 Squamous  Absent
 Transitional  Rare or few
 Renal tubular  Rare or few
ECBU

55. In-Service Training
Wetmount;
 Crystals  upto few
 Casts  Absent
Gram stain;
 Varies with wetmount
Culture;
 No growth on ‘X’ medium at T°C after ‘T’.
ECBU

56. In-Service Training
ECBU