Oleh :

Muhammad Asaad Maidin & Muh. Nasrum Massi

Bagian Mikrobiologi Fakultas Kedokteran
Universitas Hasanuddin
Makassar

ELECTRON MICROSCOPE (E.M)
COMPLEMEN FIXATION TEST
GEL DIFFUSION TEST
Polymerase Chain Reaction (PCR)
ISOLASI VIRUS
ISOLASI VIRUS ----- inokulasi pada Chorio
Allantoises membrane dari chick embrio

VARIOLA MAJOR
Small white Pocks (bercak kecil keputihan) tumbuh pada 30
o
C
ALASTRIM
Bercak kecil keputihan pocks tidak tumbuh pada 383
o
C
VACCINIA (COW-POX)
Bercak besar keabuan dgn Necrose Sentral
HERPES SIMPLEX
Bercak putih kekecilan dibanding Variola
VARICELLA
No-Pocks (tidak ada Pocks)

1. EMBRIO AYAM
Virus dapat memberikan efek sebagai berikut :
- kematian embrio yaitu virus Encefalitis
Pembentukan Pocks atau Plaque pada selaput
horioalantoik : Herpes, cacar
Pembentukan Hemaglutinasi dalam cairan atau
jaringan embrio ialah Influenza virus
Pembentukan virus infektif yaitu Polio virus tipe 2



PEMBIAKAN VIRUS

2. BIAKAN JARINGAN

- Biakan primer sanggup tumbuh hanya beberapa
kali biakan
- Biakan sekunder sampai 50 kali pemindahan
biakan
- Biakan berkesinambungan pemindahbiakan
sampai tanpa batas
Kultur jaringan untuk mengisolasi virus
( sel epitel gepeng )
Kultur Jaringan ( sel fibroblas)_
Dapat ditentukan sebagai berikut:
1. Efek sitopatik
Polio, herpes, campak, adenovirus, dsb
2. Hambatan metabolisme sel :
Entero virus
3. Timbulnya suatu Hemaglutinin Influenza
Para influenza, Influenza
4. Interferensi oleh suatu virus yang tidak sitopatogenik dalam
replikasi : - Rubella
6. Transformasi morfologik oleh virus onkogenik; SV 40



HITUNGAN VIRUS

1. Metode Fisik
- Hitung langsung : E. M
- Hemaglutinasi :
2. Sel darah manusia diaglutinasi oleh banyak
virus:
Orthomyxo virus Paramyxo virus
Pox virus Arbo virus
2. METODE BIOLOGIK

- Efek sitopatik Pengenceran suspensi virus
- L.D 50% Percobaan binatang
- Plaque Forming Unit
- Menghitung jumlah POCKS
- Khorioalantois pengenceran virus

Antivirus dan Sel yang terinfeksi
DIAGNOSIS
LABORATORY
DIAGNOSTIC OF
FUNGAL INFECTION
Examinations:
Physical examination:
predisposition
appropriate features of fungal infection
clinical history, occupation, travelling


Specimens for fungal
investigation:

swab
sputum
scrapings of skin
A small segment of infected tissues

Identifying Fungi

Yeast : biochemical tests
Filamentous :
physical appearance
spore structure
colony characteristics

Fungal investigations
involves:
Microscopic examination for evidence of hyphae or
spores
Skin scrapings in 10% KOH direct unstained smear
Sputum or ulcer swab Gram staining
Infected tissues: Direct Immunofluorescense
Infected tissues: Gomori Methenamine silver (GMS)
staining
Fungal Antigen detection
Latex agglutination test Cryptococcus as antigen

Culture media
One incubate at 37
o
C and another at room
temperature to see dimorfisme
Observe:
colony appearance: pigment, size of mycelium (varies
greatly), spore or conidia
microscopic morphology: hyphae, pseudohyphae,
mycelium. Spore, conidia, septum
Chemical reaction (carbohydrate fermentation)
Nucleic acid probe

On Sabouraud dextrose agar (acidic pH + 5,6 and
contains antibiotic)
Host response tests:
Skin tests common test, frequently false pos,
useful to evaluate host response and determine
exposure index for epidemiological purposes

Serology
Latex agglutination (detect IgM)
complement fixation (detect IgG),
frequently false pos due to cross reactions,
Antibody detection 2-3 months after onset
of disease



1. Wet Mount
2. Skin test
3. Serology
4. Fluorescent antibody
5. Biopsy and histopathology
6. Culture

Diagnosis

1. Wet Mount
2. Skin test
3. Serology
4. Fluorescent antibody
5. Biopsy and histopathology
6. Culture

Diagnosis

DIRECT MICROSCOPIC
OBSERVATION


10 % KOH

Gentle Heat
KOH Wet Mount

1. Wet Mount
2. Skin test
3. Serology
4. Fluorescent antibody
5. Biopsy and histopathology
6. Culture

Diagnosis
SKIN TESTING
(DERMAL HYPERSENSTIVITY)

Use is limited to :

Determine cellular defense
mechanisms
Epidemiologic studies


1. Wet Mount
2. Skin test
3. Serology
4. Fluorescent antibody
5. Biopsy and histopathology
6. Culture

Diagnosis
Most serological tests for fungi measure antibody.
Newer tests to measure antigen are now being
developed
ANTIGEN DETECTION PRESENTLY
AVAILABLE

Cryptococcosis
Histoplasmosis


1. Wet Mount
2. Skin test
3. Serology
4. Fluorescent antibody
5. Biopsy and histopathology
6. Culture

Diagnosis
DIRECT FLUORESCENT
ANTIBODY
CAN BE APPLIED TO
1. TISSUE
2. CULTURE

Viable
Non-viable


1. Wet Mount
2. Skin test
3. Serology
4. Fluorescent antibody
5. Biopsy and histopathology
6. Culture

Diagnosis
INFLAMMATORY
REACTION
Normal host
Pyogenic
Granulomatous
Immunodeficient host
Necrosis
Polymorphic Nuclear Leukocytes
Giant Cell
GMS

1. Wet Mount
2. Skin test
3. Serology
4. Fluorescent antibody
5. Biopsy and histopathology
6. Culture

Diagnosis
ISOLATION MEDIA
SABOURAUD DEXTROSE AGAR
(pH ~ 5.6)
Plain
With antibiotics
With cycloheximide


INCUBATION
TEMPERATURE

37 C - Body temperature

25 C - Room temperature
Microsporum
canis colony on
Sabourauds
agar
Microsporum canis macroconidia (spore)
Dermatophyte hyphae in KOH preparation of skin
scraping
Microsporium gypseum macroconidia (spore)
Histoplasma
capsulatum
macroconidia
Trichopyton rubrum colony on Sabourauds agar
Coccidioides immitis. Arthrospores after culture
Coccidioidomycosis. Lung section shows sporangiospore
Aspergillus fumigatus colony on Sabourauds agar
Cryptococcus neoformans. Budding yeast surrounded by the
large mucoid capsule is demonstrated by negative staining
with nigrosin
Aspergillus fumigatus conidiosphores
Medical Mycology Iceberg