PEMERIKSAAN

MIKROBIOLOGI
PADA PARU
Oleh :

dr. Eka Pratiwi

dr. Saidah Mafisah
 OUTLINE
01 02

PENDAHULUAN AGEN ETIOLOGI PADA

PENYAKIT PARU

03 04

PEMERIKSAAN ANTIMICROBIAL
MIKROBIOLOGI SUSCEPTIBILITY TEST

2
 PENDAHULUAN
Patogen Saluran Pernapasan Pemeriksaan Mikrobiologi

Infeksi saluran pernapasan bawah Spesimen yang paling baik dalam pengujian
disebabkan oleh berbagai macam virus dan diagnostik adalah yang dikumpulkan
bakteri patogen. Untuk pneumonia langsung dari tempat infeksi.
setidaknya, infeksi polimikrobial berurutan Ketika bakteri diisolasi seperti pada usap
atau bersamaan mungkin relatif umum tenggorokan, usap nasofaring, atau sputum,
Insiden banyak infeksi pernapasan penting mengetahui bakteri mana yang dapat
mengikuti pola siklus yang selaras dengan ditemukan sebagai komensal atau kolonisasi
transmisi musiman yang khas dari patogen di saluran pernapasan bagian atas, yang mana
tertentu. yang benar berasal dari tempat infeksi

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory illness: recent
advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405. 3
 AGEN ETIOLOGI PADA PENYAKIT PARU

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory illness: recent
advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405. 4
 AGEN ETIOLOGI PADA
PENYAKIT PARU

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory illness: recent
advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405. 5
 PEMERIKSAAN MIKROBIOLOGI

Respiratory Specimens Gram Stain Appearance

and Diagnostic Test

Molecular Terms in
Microbial Culture Diagnostic

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory illness: recent
advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405. 6
RESPIRATORY SPECIMENS & DIAGNOSTIC TESTING

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory 7
illness: recent advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405.
Respiratory Tract : Throat Swab
• Push the tongue using spatula
• Swab both tonsil and pharynx,
avoid to touch tongue or uvula

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of
respiratory illness: recent advances. Kendig's Disorders of the Respiratory Tract in
Children, pp.396-405. 8
 Respiratory Tract : Nasal swab

• Insert swab 1 cm
into the nose
• Rotate the swab, let
it 10-15 sec
• Insert to transport
medium (Amies
medium)

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of
respiratory illness: recent advances. Kendig's Disorders of the Respiratory Tract in www.thechelmsford.co.uk
Children, pp.396-405. 9
Respiratory Tract : Sputum

Sputum,
Collect sputum,
expectorated
not saliva!
sputum, inhalation

Gurgle (minimize Sterile container,

oral flora wide mouth,
contamination), screw-capped
deep coughing

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of
respiratory illness: recent advances. Kendig's Disorders of the Respiratory Tract in
Children, pp.396-405. 10
 Respiratory Tract:
Sputum for Mycobacterium

Sputum
• Sputum: (Sewaktu-Pagi/
SP)
• Acid Fast Bacilli (AFB)
staining (Ziehl Neelsen
staining)
• Sterile, wide-mouth,
screw-capped container 3-
5 ml

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of
respiratory illness: recent advances. Kendig's Disorders of the Respiratory Tract in
Children, pp.396-405. 11
 GRAM
STAINING
Pewarnaan Gram digunakan untuk
mengklasifikasikan bakteri berdasarkan
bentuk, ukuran, morfologi seluler, dan
reaksi Gram ini juga merupakan tes
penting untuk diagnosis dugaan agen
infeksi dan berfungsi untuk menilai
kualitas spesimen klinis

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological

diagnosis of respiratory illness: recent advances. Kendig's Disorders of the
Respiratory Tract in Children, pp.396-405. 12
Microscopy : Gram Staining Result

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 Microscopy: Ziehl Neelsen (ZN) staining for AFB

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory illness:
recent advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405. 14
MICROBIAL CULTURE
Sebagian besar bakteri patogen membutuhkan
24-48 jam untuk tumbuh dalam kultur, dan 24-
48 jam lagi untuk melakukan pengujian
kerentanan antimikroba.

Pasa hasil kultur bakteri saluran napas, yang

biasanya merupakan kultur campuran, analis
akan fokus pada organisme yang dominan dan
yang diketahui sebagai patogen penting  S.
Pyogenes pada usap tenggorok dan S.
pneumoniae pada sampel dahak

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory
15
illness: recent advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405.
MICROBIAL CULTURE – STREAK TECHNIQUE

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of
respiratory illness: recent advances. Kendig's Disorders of the Respiratory Tract in Children,
pp.396-405.
16
 MICROBIAL CULTURE- TYPE OF BACTERIAL COLONY

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019.

Microbiological diagnosis of respiratory illness: recent
advances. Kendig's Disorders of the Respiratory Tract in
Children, pp.396-405. 17
MOLECULAR TERMS IN
DIAGNOSTIC

Molecular microbiology:
Development of new genetically engineered
vaccines, biotechnology, antimicrobial
development, etc.

Have a direct influence on the clinical practice of

Medical Microbiology

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory illness:
recent advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405. 18
 APPLICATION OF MOLECULAR
MICROBIOLOGY
• Classification of organism (genotyping)
• Identification and confirmation of isolate obtained from
culture
• Early detection of pathogens in clinical specimen
• Rapid detection of antibiotic resistance
• Detection of mutations
• Differentiation of toxigenic from non-toxigenic strains
• For fastidious bacteria or for unculturable microbes (to
culture, grow slowly or present in extremely small numbers
in clinical specimen)

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory illness:
recent advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405. 19
 Molecular – Polymerase Chain Reaction

Murdoch, D.R., Werno, A.M. and Jennings, L.C., 2019. Microbiological diagnosis of respiratory 20
illness: recent advances. Kendig's Disorders of the Respiratory Tract in Children, pp.396-405.
MICROBES IDENTIFICATION – POLYMERASE CHAIN REACTION
Incubation
18-24 h

35 ± 20C
Blood agar plate Colonies of
bacteria

Bacterial DNA
Isolation

PCR

Gel electrophoresis

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Antimicrobial Susceptibility Tests (AST)

To predict the outcome To provide information to

of treatment with the the clinician to guide in
selecting appropriate
antimicrobial agents
antimicrobial therapy for a
tested particular clinical problem

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Routine AST Methods

Automated-instrument method –
Broth microdilution Vitek, Phoenix, Microscan,
Sensititre.

Disk Diffusion Method (Kirby-

Antimicrobial gradient (Etest,
Bauer)
MIC Evaluator)
More accessible and economic method

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AST METHODS: AUTOMATED

Results are generated in a shorter period (3.5 – 16 hours)

Computer software used to interpret AST results
Includes “expert system" for analyzing test results for atypical patterns and unusual resistance
phenotypes
Lessened ability to detect some types of antimicrobial resistance
Cost is higher
Results: MIC

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AST METHOD: GRADIENT DIFFUSION
• Commercial methods- follow manufacturer’s directions
• Same testing procedure as the disk method
• Simple and flexible, may be used to test for fastidious and anaerobic bacteria
• MICs generally agree well with MICs by standard broth or agar dilution methods
• More expensive than the disk method

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AST METHODS : DISK DIFFUSION
KIRBY - BAUER

• One of the most established and best proven of all susceptibility tests
• Continues to be updated and refined through frequent CLSI publications
• Low cost
• Results: S,I,R (based on inhibition zone interpretation)

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