Oleh :

Efrida Warganegara
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK

PENYAKIT INFEKSI

MASUK
MO
• bakteri tubuh
karena inang
• jamur patogen
• virus
• protozoa
Kelainan /
kerusakan
DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK

PENYAKIT INFEKSI

LOKAL

INFEKSI

SISTEMIK
DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK

Sehingga

Penanggulangan
Masalah penting !!!
Kesehatan
Masyarakat
• Menentukan
penyebab
• Pengobatan rasional
berdasar pem. lab
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
BAHAN PEMERIKSAAN

Penting dalam hal

Kerja sama

 Klinisi
 Paramedis
• Cara pengambilan  Ahli Lab.
• Pengiriman  Ahli
Mikrobiologi
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK

DIAGNOSIS

HASIL
PEMERIKSAAN DIAGNOSA KLINIK
MIKROBIOLOGI
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
KEGAGALAN PEMERIKSAAN MIKROBIOLOGI

Tidak berarti
Sebab
kegagalan
• Di Lab : teknik dan
carakerja
• Cara pengambilan
Diagnosis klinik
dan pengiriman BP
salah
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK

PERAN AHLI MIKROBIOLOGI KLINIK

* Penetapan penyebab infeksi

* Penentuan terapi antimikroba
yang sesuai
* Penatalaksanaan infeksi
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
PERAN AHLI MIKROBIOLOGI KLINIK

PENTING ???

Hasil pemeriksaan negatif

Tidak
menyingkirkan
Yang dihadapi
?
bukan suatu penyakit infeksi
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
PATOKAN UMUM PENGAMBILAN BAHAN :
1. JUMLAH BAHAN CUKUP

2. REPRESENTATIF
3. STERILITAS ALAT & CARA-CARA
ASEPTIK
4. STADIUM
PENYAKIT
5. SEBELUM Th /
ANTIBIOTIK
6. SEGERA DIBAWA KE
LABORATORIUM
dan SEGERA DIPERIKSA
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
PATOKAN UMUM PENGAMBILAN BAHAN :

7. KETERANGAN KLINIK

• NAMA, UMUR,JENIS
• KELAMIN
RS / DOKTER / BAG.
YANG MENGIRIM
• TANGGAL
PENGAMBILAN B.P.
• JENIS BAHAN
PEMERIKSAAN
• KETERANGAN KLINIK
PENDERITA
• PEMERIKSAAN YANG
DIMINTA
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
INFEKSI ANAEROB
SIFAT-SIFAT :
• Sering berhubungan dengan permukaan
mukosa
• Cenderung m.o. campuran
• Cenderung infeksi tertutup  abses
( dalam )
• Pus bau busuk
• O.k. suplai darah menurun  jaringan
nekrotik  tegangan o2 menurun
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
INFEKSI ANAEROB

BAHAN PEMERIKSAAN ANAEROB

* Cara pengambilan : anaerob

* Medium transpor khusus

* Secepatnya ke Laboratorium
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK

PENGIRIMAN BP MEDIUM
TRANSPORT
Menjaga agar m.o
tetap tumbuh / hidup
 Tempat tertutup
• M.o. tersangka mampu
• Es kering ± 40C hidup
• M.o. lain tidak tumbuh
• Label
berlebihan (tidak
menekan m.o. yang
dicari)
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
CARA PENGAMBILAN DAN PENGIRIMAN BP
1. DARAH.
TANPA MEDIUM
* PUNKSI VENA,
TINDAKAN ASEPTIK
• VOL DARAH : VOL. DENGAN
MEDIUM ANTIKOAGULAN
HARUS SEGERA
1 : 5• MEDIUM:
atau 1 : 10
DIKIRIM !
– Tioglikolat Jangan lakukan palpasi
– Tioglikolat Brain setelah tindakan
– Heart Infusion Broth ANTISEPSIS
(BHI)
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
CARA PENGAMBILAN DAN PENGIRIMAN BP

2. URINE
• Aspirasi suprapubis Vol. Urine minimal : 10 cc
• Kateterisasi ( 3 jam tidak berkemih )

DILAKUKAN
DOKTER
• Tengah arus
Urine :
• Alat genital dicuci :
TANPA MEDIUM TRANSPORT
– sabun HARUS DITANAM < 1 JAM
– air bersih ( 24 JAM, LEMARI ES )
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
CARA PENGAMBILAN DAN PENGIRIMAN BP

3. Dahak / sputum
Batuk dalam
Tanpa medium
transport
Segera kirim

• Sputum pagi
• Sputum sewaktu

± 1 - 3 ml
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
CARA PENGAMBILAN DAN PENGIRIMAN BP

4. TIN JA
Bahan Pemeriksaan diperoleh
dari :
• Apus rektum
• Tinja
1. Tanpa medium 2. Dengan medium
transport transport :
* Air pepton alkali
* Selenit Broth
DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
CARA PENGAMBILAN DAN PENGIRIMAN BP

5. SELAPUT LENDIR / KULIT

Swab dari selaput lendir :

o Hidung 0 Tenggorok
o Mata o Telinga
o Lubang urogeniotal o
Luka
Medium Transport :
• Stuart
• Carry Blair
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
CARA PENGAMBILAN DAN PENGIRIMAN BP
6. CAIRAN CEREBROSPINALIS

Punksi
Punksi lumbal Lumbal
Jangan disimpa
dlm lemari es
• Sterilitas alat
• Aseptik
Sumber : Color Atlas of Medicine and Parasitology. 1977
W. Peters & H.M. Gillers
Tanpa medium transport
BEDSIDE CULTURE
Kirim < 1 jam
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
CARA PENGAMBILAN DAN PENGIRIMAN BP

7. LUKA, JARINGAN, TULANG, ABSES, & CAIRAN TUBUH

LAIN

• Biopsi jaringan
• Pus aspirasi
abses
• Eksudat
– rongga pleura
– rongga cinovia dll
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
TEKNIK-TEKNIK PEMERIKSAAN MIKROBIOLOGI

1. PEMERIKSAAN LANGSUNG 
PEWARNAAN
*
Penyarin
* Pengobatan awal sebelum hasil
g
biakan
Pewarnaan
* Neisser
* Ziehl-Neelsen Bakteri Tahan Asam
( BTA)
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
TEKNIK-TEKNIK PEMERIKSAAN MIKROBIOLOGI
1). PEMERIKSAAN LANGSUNG  PEWARNAAN
* Pd bbp infeksi ttt cara ini sangat spesifik dan sensitif  dpt
segera diterapi tanpa dikultur dulu
* menghasilkan 3 informasi :
1. Konfirmasi adanya m.o. pd cairan tubuh yg steril
2. Sifat dan morfologi m.o.
3. menghasilkan diagnostik klinik
* Misal : - Infeksi cacing, protozoa
- Jamur patogen : kapsul (+) pd Cryptocococcus
neoformans
- Sifilis : Spirochaeta (bentuk & gerakan)
- Gonorrhoeae : m.o. Gram (-), diplokokus,
intraseluler leukosit
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
TEKNIK-TEKNIK PEMERIKSAAN MIKROBIOLOGI

2). ISOLASI
IDENTIFIKASI

• KONVENSION • OTOMATISASI
AL
3). UJI KEPEKAAN terhadap
Antibiotika
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
TEKNIK-TEKNIK PEMERIKSAAN MIKROBIOLOGI

4). SEROLOGI :

• Presipitasi • Imunofluorese
• Aglutinasi nsi
• Reaksi • Elisa
ikatan • RIA
komplemen • LIA
•
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
TEKNIK-TEKNIK PEMERIKSAAN MIKROBIOLOGI

5). TEKNIK BIOLOGI

MOLEKULER :

• Polimerase Chain Reaction

(PCR)
• Pelacak DNA
DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
UJI SEROLOGI

Imunodiagnost
• Deteksi Antigenik
( Ag ) atau Antibodi
( Ab )
• Atau
DASAR : Ab
Ag dan
REAKSI Ag - Ab
Tes imunodiagnostik
Bahan pemeriksaan bermacam-macam:
Sputum, Tinja, Sekret urogenital dll.
Serologi
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
UJI SEROLOGI
1. Presipitasi :
- Ag dan Ab dlm btk larutan
- ada 2 macam reaksi (presipitasi dlm cairan dan
presipitasi btk pita, cakram, garis dlm media agar
= difusi)
- misal : Test VDRL -> lues

2. Aglutinasi :
- Ag suspensi, dan Ab larutan
- misal : Test Widal -> Typhus abdominalis
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
UJI SEROLOGI : (lanjutan)

3. Reaksi Ikatan Komplemen

- memeriksa Ab yg dpt mengikat komplemen secara tdk
langsung dgn menambahkan sistem hemolitik
- (+) : bila komplemen digunakan oleh Ab, maka reaksi
hemolitik tidak terjadi
(-) : bila komplemen tidak digunakan oleh Ab, reaksi
hemolitik akan terjadi.
- misal : Test Wasserman -> Lues

4. Tes Netralisasi
- Dengan menambahkan Ab spesifik dari virus kedalam biakan
virus yg sedang aktif metabolismenya  maka aktivitas
metabolisme tsb akan dinetralkan
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
UJI SEROLOGI : (lanjutan)
5. Imunofluoresensi
- Ab dikonjugasikan dgn bahan fluoresens -> campur dgn
sampel -> lalu mengikat Ag-nya. Ag yg terikat Ab ->
dapat dilihat dibawah mikroskop fluoresensi

6. Elisa (Enzym Linkage Sorbent Assay)

- Ab dikonjugasikan dgn enzim -> campur dgn sampel ->
lalu mengikat Ag-nya. Ag yg terikat Ab -> dapat diukur dgn
spektrofotometer
- Enzimnya yi : peroksidase, Alkalis fosfatase, Lisozym

7. RIA (Radio Imun Assay)

 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
TUJUAN PERBENIHAN KUMAN

UNTUK PEMBIAKAN

MEMPERBANYAK
• ISOLASI BIAKAN
MURNI
• PENYIMPANAN KUMAN MURNI
• MEMPELAJARI SIFAT-SIFAT
PERTUMBUHAN
DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
IDENTIFIKASI KUMAN

• KOKUS GRAM
POSITIF

• KOKUS GRAM
NEGATIF

• BATANG GRAM
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
MACAM-MACAM PERBENIHAN

Berdasar sifat kuman terhadap

hospes :
• Medium hidup
• Medium
Mediummati
Hidup :
• Hewan percobaan
• Telur ayam berembrio
• Biakan jaringan
Sel-sel biakan bakteri
 DASAR-DASAR MIKROBIOLOGI
KLINIK DIAGNOSTIK
MACAM-MACAM PERBENIHAN
Berdasar sifat kuman terhadap
hospes :
• Medium hidup
•Medium
MediumMati
mati

1. Konsistensi 2. Fisiobiologik
• Medium
• Padat : Petri, Tabung persemaian
• Semisolid : Gerak • Medium
• Cair : Reaksi Biokimia, Uji diperkaya
DIAGNOSTIC METHODS
OF VIRAL INFECTION
DIAGNOSTIC METHODS OF VIRAL
INFECTION
1. Clinical symptoms
2. Laboratory diagnosis

Typical clinical symptoms

- Polyomyelitis
- Chicken Pox
- Measles
- Mumps
Laboratory diagnosis
1. Electron microscope morphology
2. Light microscope special staining
~ type of virus
 Variola virus inclusion bodies
Gispen staining : PASCHEN BODIES
 Rabies virus specimen : brain
Sellers staining : inclusion bodies in nerve
cells NEGRI BODIES
 Molluscum contagiosum virus skin nodule
Lugol staining : inclusion bodies in
cytoplasm of epithel cell
MOLLUSCUM
BODIES
Laboratory diagnosis :
3. Culture
specimen : depend on the disease
in vitro, in ovo, or in vivo
4. Serology
 Raise of antibody titer
 Antigen detection from the specimen
 Viral type identification :
agglutination, precipitation, complement
fixation test, neutralization, inhibition
haemagglutination, FAT, ELISA, RIA, RIA
DIAGNOSTIC METHODS
OF FUNGAL INFECTION
 BASIC LABORATORY
PROCEDURES
Successful isolation of a fungus causing deseases is
dependent upon each of the following factors :
 proper collection of specimen
 proper handling and correct processing of the
specimen (including the inoculation of the specimen
onto the appropriate culture medium and incubation at
a suitable temparature
 the expertise of the technologist for identifying the
fungus
Collection of specimens :

1. Skin scrapings :
 clean the lesion of dirt or any topical medicines
 scrape the outer edge of the lesion with a scalple
 collect the scrapings in a clean container

2. Hair :  remove hair from the infected site with clean forceps
 collect in a clean container

3. Biopsied tissue :
 placed in a sterile containers;
 add steril water or saline to keep the tissue moist
 do not freeze the tissue

4. Exudate or pus :
 should be aspirated from an unopened abscess
 placed in a steril tube and taken directly to the laboratory
 never let the specimen dry
Collection of specimens :
5. Sputum :
 collect sputum early in the morning as soon as the
patient awakens
 before collecting the sputum, the patient should
brush his teeth or remove his dentures, then thoroughly
rinse his mouth
 ask the patient to take a deep cough and raise sputum
from the lung
 collect the sputum in a wide-mouthed, sterile container
that can be tightly closed to prevent leakage
 send the specimen directly to the laboratory
Collection of specimens :

6. Blood :  should be collected aseptically

 placed in the culture medium at the
patient’s bedside

7. Spinal fluid :
 collect aseptically and place into a sterile
tube
 do not refrigerate spinal fluid

8. Urine :  collect in asterile container

 take directly to the laboratory
Processing of specimens :

Specimens should be processed as soon as possible :

 to ensure that the infecting fungus does not die
 to control contaminating organism

If the specimen cannot be promtly processed, it should be

refrigerated (except spinal fluid).

1. Sputum and bronchial washings :

 examine specimen grossly for purulent or caseous
material or particles
 prepare smears and wet mounts and inoculate this
material onto appropriate culture media
Processing of specimens :
2. Spinal fluid, urine and pleural fluid :
 concentrate the specimen by centrifugation
 make a wet preparation of the sediment and inoculate
the appropriate media with the remaining sediment
3. Tissue taken by surgical procedure :
 remove any caseous or purulent material and place
onto the appropriate media & prepare wet preparation &
smears
 cut the tissue into small pieces with sterile scissors
and grind the tissue with a sterile mortar and pestle
 transfer the homogenized tissue to appropriate
media
Processing of specimens :
Direct microscopic examination :
 is an essential step in diagnosing a fungal
disease
 often provide a rapid, tentative diagnosis (without
having to wait for the
culture to grow)
 culture must always be made to correctly identify
the fungus
 most mycological specimens are examined in
the fluid state (wet mount), include a KOH (or
NaOH) preparation, India ink. lactophenol- cotton
blue
Processing of specimens :
Culture media for isolation and identification :
The proper selection of isolation media is critical to obtaining a
laboratory diagnosis of a fungal disease. If the wrong medium
is used, the fungus causing disease may not grow.
Culture media routinely used may be divided into two
main
 primary isolation media (non-selective or selective -- ->
may contain antibiotics to inhibit rapidly growing fungi)
 differential media; are used to identify selected genera or
or species (by stimulation of characteristic
growth/sporulation; or by the production of physiological
reaction on these media)
Processing of specimens :
Culture media for isolation and identification :
 the isolation medium selection depends upon :
 the type of specimen (heavily contaminated,
or sterile)
 the suspected etiological agent
A non selective medium like Sabouraud dextrose
agar (SDA) should be routinely used because it
will support the growth of almost all the
medically important fungi.
However, without the addition of selective agent
(such as chloramphenicol and cycloheximide) this
medium is practically useless.
TERIMA
KASIH
PREVENTION AND TREATMENT OF VIRAL
INFECTIONS
1. VIRAL VACCINES
 Killed-virus vaccines
 Attenuated live-virus vaccines
 Future prospect :
- attenuation of viruses by genetic
mapping
- avirulent viral vectors
- purified proteins produced using
cloned genes
- synthetic peptides
- subunit vaccines
- DNA vaccines
2. INTERFERONS
IFNs :
 host-coded proteins of large cytokine
family
 inhibit viral replication
 produced by intact animal or cell
culture in response to viral infection
or other inducers
 first line of defense against viral
infection
 3. ANTIVIRAL CHEMOTHERAPY

A. Nucleoside analogs
 Acyclovir &  Ribavirin
valacyclovir Stavudine (d4T)
 Didanosine Trifluridine
 Gancyclovir Vidarabine
 Idoxuridine Zalzitabine (ddC)
 Lamivudin (3TC) Zidovudine (AZT)
B. Nucleotide analogs
Cidofovir : active against CMV & HSV
inhibit s viral DNA polymerase
C. Nonnucleoside reverse transcriptase inhibitor
Nevirapine : inhibit reverse transcriptase of HIV
D. Protease inhibitors
Ritonavir, Saquinavir HIV
E. Other types
Amantadine & rimantadine
Foscarnet
Methiasone