Direct Sputum Smear Microscopy (DSSM) PDF

DIRECT SPUTUM SMEAR
MICROSCOPY (DSSM)/ AFB STAIN
JON PATRICK MINA, RMT

BOW OUR HEADS AND FEEL THE PRESENCE
OF THE LORD…
LEARNING OBJECTIVES:
• AT THE END OF THIS SESSION, THE READER MUST BE

ABLE TO:
• 1. DISCUSS THE IMPORTANCE OF THIS PROCEDURE TO THE
LABORATORY PROFESSION
• 2. DISCUSS THE STEPS OF DSSM
• 3. CORELATE THE RESULT OF DSSM TO DIAGNOSIS OF THE
PATIENT
• 4. DISCUSS THE REPORTING OF THE RESULT IN TERMS OF
DIRECT MICROSOPY
BACKGROUND
• Worldwide, TB is the 13th leading cause

of death and the second leading
infectious killer after COVID-19 (above
HIV/AIDS).
• total of 1.5 million people died from TB in
2020 (including 214 000 people with HIV).
• In 2020, an estimated 10 million people
fell ill with tuberculosis (TB) worldwide.
• 5.6 million men
• 3.3 million women and
• 1.1 million children
IN 2020…
• There is 30 high TB burden countries

accounted for 86% of new TB cases
• Eight countries account for two thirds of
the total population
• India
• China,
• Indonesia
• Philippines,
• Pakistan,
• Nigeria,
• Bangladesh and
• South Africa
PHILIPPINES TUBERCULOSIS ROADMAP
OVERVIEW, FISCAL YEAR 2021
• Estimated to have a 599,000
active tb case
• Ranked as seventh in the MDR
TB burden countries
Tuberculosis (TB)
• is caused by a bacterium called Mycobacterium
tuberculosis.
• The bacteria usually attack the lungs
• Kidney
• spine, and
• brain
• two TB-related conditions exist:
• latent TB infection (LTBI) and
• TB disease
Diagnosis of TB Disease
• Diagnosed By
• medical history,
• physical examination,
• chest x-ray, and
• other laboratory tests
• TB disease is treated by taking several

drugs as recommended by a health
care provider.
LABORATORY IDENTIFICATION
OF TUBERCULOSIS
DSSM /AFB
STAIN
Collection and transport of sputum
specimens
• Instruct patient to expectorate sputum sample
• 2 TYPES OF SAMPLE
• 1. SPOT-SPOT
• 2. EARLY MORNING – SPOT
• SM/TB LAMP/XPERT
• XPERT MTB/RIF / LAMP
• SMEAR MICROSCOPY
• Collect specimen in a well-ventilated designated sputum
collection area, or outside the health facility
• Instruct the patient on how to expectorate:

• a. clean mouth by thoroughly rinsing with water
• b. breathe deeply, hold breath for a second or two, and then
exhale slowly. Repeat the entire sequence two more times;
• c. cough strongly after inhaling deeply for the third time and try
to bring up sputum from deep within the lungs; and
• d. expectorate the sputum in the sputum cup or conical tube
ASSESMENT OF QUALITY OF
SPUTUM
Notes to remember
• For SM, instruct to collect a second sample one hour later

or an early-morning sputum sample the following day.
• Follow-up within three days if patient fails to submit a

second specimen unless the first specimen already tests
positive for acid-fast bacillus (AFB) in which case the
second specimen will not be necessary.
COILING OF SPUTUM SAMPLE
3 IMPORTANT
PARAMETERS TO
CONSIDER:
1. THICKNESS
2. EVENESS
3. SIZE
ACID FAST STAIN
• - It is used to stain bacteria that have high lipid content in their
cell walls.
• Principle:
• The primary stain binds to mycolic acid in the cell walls of the
mycobacteria and is retained after decolorozation with acid
alcohol.
• Acid-fast bacilli (AFB) retain the primary stain and are colored
red (carbol fuschin) and
• non-AFB blue or green (methylene blue or malachite green
counterstains) color. (resistant to decolorization)
• Non-Acid fast don’t contain mycolic acid and become
decolorized with alcohol
ACID FAST STAINING METHODS
• 1. Ziehl-Neelsen/Hot Staining Method – uses steam (physical

mordant)
• 2. Kinyoun’s/Cold Staining Method – same reagent but omits
heat (carbol fuchsin has higher concentration of phenol which
serves as a chemical mordant); sometimes have a wetting
agent tergitol to fasten up reaction
• 3. Pappenheim’s Method – differentiates M. smegmatis from M.
tuberculosis
• 4. Baumgarten’s Method – differentiates M. leprae from M.
tuberculosis
• 5. Auramine-Rhodamine Method – selective for the cell wall of
AFB
PROPER WAY OF READING
L
A
B
E
L
150 QVF
• THANK YOU!!!
• PREPARE FOR QUIZ NEXT MEETING.

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DIRECT SPUTUM SMEAR

MICROSCOPY (DSSM)/ AFB STAIN

JON PATRICK MINA, RMT

• AT THE END OF THIS SESSION, THE READER MUST BE

• Worldwide, TB is the 13th leading cause

• There is 30 high TB burden countries

• TB disease is treated by taking several

• Instruct the patient on how to expectorate:

• For SM, instruct to collect a second sample one hour later

• Follow-up within three days if patient fails to submit a

• 1. Ziehl-Neelsen/Hot Staining Method – uses steam (physical

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