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SPUTUM
EXAMINATION
PRESENTED BY:
 MARTEY ALFRED
OBJECTIVES
WHAT IS SPUTUM
PHYSICAL DESCRIPTION OF A GOOD SPUTUM
SPUTUM COLLECTION PROCEDURE
SPUTUM AFB
STANDARD OPERATION PROCEDURE FOR SPUTUM
SMEAR MICROSCOPY
COMMON PROCEDURES FOR SPUTUM SMEAR
MICROSCOPY
AFB STAINING TECHNIQUES USING THE FLOURESCENT
METHOD
8/21/2015 Fenteng et al 2015 KOGH 5 2
SPUTUM
Sputum is mucus that is coughed up from the lower
airways. This process is known as sputilization. In
medicine, sputum samples are usually used for
microbiological investigations of respiratory infections
and cytological investigations of respiratory systems.
8/21/2015 Fenteng et al 2015 KOGH 5 3
8/21/2015 Fenteng et al 2015 KOGH 5 4
Physical description of a good sputum
Purulent sputum contains pus,
composed of white blood cells,
cellular debris, dead tissue, serous
fluid, and viscous liquid (mucus).
Purulent sputum is typically yellow
or green. It is seen in cases of
bronchitis or acute upper
respiratory tract infection
(common cold, laryngitis).
8/21/2015 Fenteng et al 2015 KOGH 5 5
A bacterial sputum culture is used to detect and
diagnose bacterial lower respiratory tract infections
such as bacterial pneumonia, Emphysemia and
bronchitis. It is typically performed with a Gram stain to
identify the bacteria causing a person's
infection(Sabatine, 2014).
8/21/2015 Fenteng et al 2015 KOGH 5 6
8/21/2015 Fenteng et al 2015 KOGH 5 7
When is it ordered?
A bacterial sputum culture is ordered when a health practitioner
suspects that someone has a bacterial infection of the lungs or
airways(American Association of Clinical Chemistry, 2014).
Symptoms may include:
Cough
Fever
Muscle aches
Fatigue
Trouble breathing
Chest Pain
8/21/2015 Fenteng et al 2015 KOGH 5 8
What does the test result mean?
Microbiological sputum samples are usually used to look
for infections by Moraxella catarrhalis, Mycobacterium
tuberculosis, Streptococcus pneumoniae, and
Haemophilus influenza.
8/21/2015 Fenteng et al 2015 KOGH 5 Streptococcus pneumoniae9
The most common cause of bacterial pneumonia
in adults in the U.S. is Streptococcus pneumoniae
(pneumococcus)(Kämpe, 2013). Other common bacteria
include:
Staphylococcus aureus (staph)
Haemophilus influenzae
Moraxella catarrhalis
Klebsiella pneumoniae
Streptococcus pyogenes
8/21/2015 Fenteng et al 2015 KOGH 5 10
8/21/2015 Fenteng et al 2015 KOGH 5 11
8/21/2015 Fenteng et al 2015 KOGH 5 12
Standard operating procedures for
sputum smear microscopy
Numbering on a slide
Smearing sputum on the slide
1. Break a wooden applicator into two pieces to make one end
rough
Length of the stick should be at least 10cm
2. Pick sputum from the most muco-purulent part of the wooden
applicator
8/21/2015 Fenteng et al 2015 KOGH 5 13
3. Gently spread the sputum in the centre of the slide in a uniform
manner to get the size between 1x2cm and 2x3cm circular shape.
4. Allow the smear to dry completely
8/21/2015 Fenteng et al 2015 KOGH 5 14
 Fixing the smears
•With a forceps, hold the slide and pass it over a flame
2-3times
• Do not over heat or burn the smear
8/21/2015 Fenteng et al 2015 KOGH 5 15
SOME COMMON METHODS FOR SPUTUM SMEAR
MICROSCOPY
Complete Blood Count
AFB staining techniques
•Ziehl-Neelsen Method
•Fluorescence Method
Bacterial sputum culture
8/21/2015 Fenteng et al 2015 KOGH 5 16
PROCEDURE USING FLUORESCENT METHOD
AURAMINE STAINY OF SLIDE
1. Arrange slide in serial order on stainy bridge with the
smear up
8/21/2015 Fenteng et al 2015 KOGH 5 17
2. Apply filtered 0.1% Auramine on the slide and
keep for at least 25minutes
8/21/2015 Fenteng et al 2015 KOGH 5 18
3. Rinse with water and drain.
8/21/2015 Fenteng et al 2015 KOGH 5 19
4. Apply 0.5% HCL decolorizing solution for 3 minutes
8/21/2015 Fenteng et al 2015 KOGH 5 20
5. Rinse with water and drain
8/21/2015 Fenteng et al 2015 KOGH 5 21
6. Apply 0.5% potassium permanganate for 1 minute
8/21/2015 Fenteng et al 2015 KOGH 5 22
7. Rinse with water and drain
8. Air dry on slide rack.
8/21/2015 Fenteng et al 2015 KOGH 5 23
Reporting format
X200 Magnification
(1line=2cm=30fields)
>100 AFB/ field on average (3+)
10-100 AFB/ field on average (2+)
30-299 AFB/line (1+)
1-19 AFB/line (scanty)
No AFB found/line (-)
(LeBlond, 2007)
EXAMINE
FLUORESCENT MICROSCOPE8/21/2015 Fenteng et al 2015 KOGH 5 24
X400 magnification
(1line=2cm=40fields)
>50 AFB/ field on average (3+)
5-50 AFB/ field on average (2+)
20-199 AFB/line (1+)
1-19 AFB/line (scanty)
No AFB found/line (-)
8/21/2015 Fenteng et al 2015 KOGH 5 25
CONCLUSION
Sputum is not sterile so when a person has an infection,
there will typically be pathogenic bacteria present.
Specific techniques are carried out to identify such
bacteria then antimicrobial susceptibility testing is
usually performed so that the appropriate antibiotics
can be prescribed(AACC, 2014).
8/21/2015 Fenteng et al 2015 KOGH 5 26
8/21/2015 Fenteng et al 2015 KOGH 5 27
REFERENCES
1. Richard F.LeBlond. Diagnostics_expectoration. US: McGraw-Hill Companies, Inc.
ISBN 0-07-140923-8.
2. Richard F.LeBlond. Diagnostics_expectoration. US: McGraw-Hill Companies, Inc.
ISBN 0-07-140923-8
3. Richard F.LeBlond. Diagnostics. US: McGraw-Hill Companies, Inc. ISBN 0-07-
140923-8.
4. Uppsala Academic Hospital > Guidelines for treatment of acute lung diseases.
August 2004. Authors: Christer Hanson, Carl-Axel Karlsson, Mary Kämpe, Kristina
Lamberg, Eva Lindberg, Lavinia Machado Boman, Gunnemar Stålenheim
5. Sabatine, [edited by] Marc S. (2014). Pocket medicine (Fifth edition. ed.). [S.l.]:
Aspen Publishers, Inc. ISBN 1451193785
6. American Association of Clinical Chemistry, bing.com/AACC
8/21/2015 Fenteng et al 2015 KOGH 5 28

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Sputum Examination and Analysis

  • 2. OBJECTIVES WHAT IS SPUTUM PHYSICAL DESCRIPTION OF A GOOD SPUTUM SPUTUM COLLECTION PROCEDURE SPUTUM AFB STANDARD OPERATION PROCEDURE FOR SPUTUM SMEAR MICROSCOPY COMMON PROCEDURES FOR SPUTUM SMEAR MICROSCOPY AFB STAINING TECHNIQUES USING THE FLOURESCENT METHOD 8/21/2015 Fenteng et al 2015 KOGH 5 2
  • 3. SPUTUM Sputum is mucus that is coughed up from the lower airways. This process is known as sputilization. In medicine, sputum samples are usually used for microbiological investigations of respiratory infections and cytological investigations of respiratory systems. 8/21/2015 Fenteng et al 2015 KOGH 5 3
  • 4. 8/21/2015 Fenteng et al 2015 KOGH 5 4
  • 5. Physical description of a good sputum Purulent sputum contains pus, composed of white blood cells, cellular debris, dead tissue, serous fluid, and viscous liquid (mucus). Purulent sputum is typically yellow or green. It is seen in cases of bronchitis or acute upper respiratory tract infection (common cold, laryngitis). 8/21/2015 Fenteng et al 2015 KOGH 5 5
  • 6. A bacterial sputum culture is used to detect and diagnose bacterial lower respiratory tract infections such as bacterial pneumonia, Emphysemia and bronchitis. It is typically performed with a Gram stain to identify the bacteria causing a person's infection(Sabatine, 2014). 8/21/2015 Fenteng et al 2015 KOGH 5 6
  • 7. 8/21/2015 Fenteng et al 2015 KOGH 5 7
  • 8. When is it ordered? A bacterial sputum culture is ordered when a health practitioner suspects that someone has a bacterial infection of the lungs or airways(American Association of Clinical Chemistry, 2014). Symptoms may include: Cough Fever Muscle aches Fatigue Trouble breathing Chest Pain 8/21/2015 Fenteng et al 2015 KOGH 5 8
  • 9. What does the test result mean? Microbiological sputum samples are usually used to look for infections by Moraxella catarrhalis, Mycobacterium tuberculosis, Streptococcus pneumoniae, and Haemophilus influenza. 8/21/2015 Fenteng et al 2015 KOGH 5 Streptococcus pneumoniae9
  • 10. The most common cause of bacterial pneumonia in adults in the U.S. is Streptococcus pneumoniae (pneumococcus)(Kämpe, 2013). Other common bacteria include: Staphylococcus aureus (staph) Haemophilus influenzae Moraxella catarrhalis Klebsiella pneumoniae Streptococcus pyogenes 8/21/2015 Fenteng et al 2015 KOGH 5 10
  • 11. 8/21/2015 Fenteng et al 2015 KOGH 5 11
  • 12. 8/21/2015 Fenteng et al 2015 KOGH 5 12
  • 13. Standard operating procedures for sputum smear microscopy Numbering on a slide Smearing sputum on the slide 1. Break a wooden applicator into two pieces to make one end rough Length of the stick should be at least 10cm 2. Pick sputum from the most muco-purulent part of the wooden applicator 8/21/2015 Fenteng et al 2015 KOGH 5 13
  • 14. 3. Gently spread the sputum in the centre of the slide in a uniform manner to get the size between 1x2cm and 2x3cm circular shape. 4. Allow the smear to dry completely 8/21/2015 Fenteng et al 2015 KOGH 5 14
  • 15.  Fixing the smears •With a forceps, hold the slide and pass it over a flame 2-3times • Do not over heat or burn the smear 8/21/2015 Fenteng et al 2015 KOGH 5 15
  • 16. SOME COMMON METHODS FOR SPUTUM SMEAR MICROSCOPY Complete Blood Count AFB staining techniques •Ziehl-Neelsen Method •Fluorescence Method Bacterial sputum culture 8/21/2015 Fenteng et al 2015 KOGH 5 16
  • 17. PROCEDURE USING FLUORESCENT METHOD AURAMINE STAINY OF SLIDE 1. Arrange slide in serial order on stainy bridge with the smear up 8/21/2015 Fenteng et al 2015 KOGH 5 17
  • 18. 2. Apply filtered 0.1% Auramine on the slide and keep for at least 25minutes 8/21/2015 Fenteng et al 2015 KOGH 5 18
  • 19. 3. Rinse with water and drain. 8/21/2015 Fenteng et al 2015 KOGH 5 19
  • 20. 4. Apply 0.5% HCL decolorizing solution for 3 minutes 8/21/2015 Fenteng et al 2015 KOGH 5 20
  • 21. 5. Rinse with water and drain 8/21/2015 Fenteng et al 2015 KOGH 5 21
  • 22. 6. Apply 0.5% potassium permanganate for 1 minute 8/21/2015 Fenteng et al 2015 KOGH 5 22
  • 23. 7. Rinse with water and drain 8. Air dry on slide rack. 8/21/2015 Fenteng et al 2015 KOGH 5 23
  • 24. Reporting format X200 Magnification (1line=2cm=30fields) >100 AFB/ field on average (3+) 10-100 AFB/ field on average (2+) 30-299 AFB/line (1+) 1-19 AFB/line (scanty) No AFB found/line (-) (LeBlond, 2007) EXAMINE FLUORESCENT MICROSCOPE8/21/2015 Fenteng et al 2015 KOGH 5 24
  • 25. X400 magnification (1line=2cm=40fields) >50 AFB/ field on average (3+) 5-50 AFB/ field on average (2+) 20-199 AFB/line (1+) 1-19 AFB/line (scanty) No AFB found/line (-) 8/21/2015 Fenteng et al 2015 KOGH 5 25
  • 26. CONCLUSION Sputum is not sterile so when a person has an infection, there will typically be pathogenic bacteria present. Specific techniques are carried out to identify such bacteria then antimicrobial susceptibility testing is usually performed so that the appropriate antibiotics can be prescribed(AACC, 2014). 8/21/2015 Fenteng et al 2015 KOGH 5 26
  • 27. 8/21/2015 Fenteng et al 2015 KOGH 5 27
  • 28. REFERENCES 1. Richard F.LeBlond. Diagnostics_expectoration. US: McGraw-Hill Companies, Inc. ISBN 0-07-140923-8. 2. Richard F.LeBlond. Diagnostics_expectoration. US: McGraw-Hill Companies, Inc. ISBN 0-07-140923-8 3. Richard F.LeBlond. Diagnostics. US: McGraw-Hill Companies, Inc. ISBN 0-07- 140923-8. 4. Uppsala Academic Hospital > Guidelines for treatment of acute lung diseases. August 2004. Authors: Christer Hanson, Carl-Axel Karlsson, Mary Kämpe, Kristina Lamberg, Eva Lindberg, Lavinia Machado Boman, Gunnemar Stålenheim 5. Sabatine, [edited by] Marc S. (2014). Pocket medicine (Fifth edition. ed.). [S.l.]: Aspen Publishers, Inc. ISBN 1451193785 6. American Association of Clinical Chemistry, bing.com/AACC 8/21/2015 Fenteng et al 2015 KOGH 5 28