Anda di halaman 1dari 38

DIAGNOSTIK PENYAKIT

INFEKSIUS

Fedik A. Rantam, DVM., Dr., Prof.


Lab. Virologi dan Imunologi, FKH UA; Stem Cell Research and
Development Center (SCDRC) UA
Surabaya
CENTRAL DOGMA
CENTRAL DOGMA DNA/RNA
-3, -2, -1 0 +1, +2, +3
DNA/RNA ASAM SYMPTOME
PEPTIDA POLIPEPTIDA PROTEIN
AMINO

Genom Protein
1. Konvensional 1. ELISA
PCR 2. IFA
2. Multiplex PCR 3. AGPT
3. Real time PCR 4. CFT
4. Droplet PCR 5. Western
5. NASBA blot
6. Micro Array 6. Strip
7. Insitu 7. SN-Test
Hybridization 8. VN-Test
ENZYME LINKAGE
IMMUNOABSORBANCE
ASSAY (ELISA)
Fedik A. Rantam, DVM., Dr. Prof.
Lab. Virologi dan Imunologi, FKH UA; Stem Cell Research and
Development Center (SCDRC) UA
Surabaya
ELISA
• Enzyme-Linked Immunosorbent Assay (ELISA), used to detect
a target substance in a liquid sample
• is a frequently used application which determines the concentration
of an analyte in solution
Macam ELISA
Tahapan ELISA
LATEX ELISA
Prinsip Latex ELISA
Latex ELISA
Latex Nanoparticle
Protein Strip
Purifikasi Protein
SEL ELISA
SEL ELISA
Bahan: Kultur sel pada mikroplate, aceton-20oC,
PBS-T 0,05%, BSA 1%, Antibodi, Konjugate
fragment Fab anti IgG-HRP-
peroxidase/alkaline phosphatase, Substrat
OPD/NPP, HCl 1N/NaOH
Keuntungan: tidak diperlukan coating antigen
selama 24 h
Kerugiannya: hanya untuk deteksi antigen
PROSEDUR SEL ELISA
1. Sel yang diinfeksi dengan virus
difiksasi dengan aceton -20oC selama
15 men
2. Dicuci 3 kali dengan PBS-T 0,05%
3. Dibloking dengan BSA 1%
4. Cuci seperti ad 2
5. Reaksikan dengan antibodi spesifik
dan inkubasikan selama 45 men pada
temp 37oC
6. Cuci seperti ad 2
7. Tambahkan konjugate
peroksidase/alkaline phosphatase dan
inkubasikan pada temp 37oC selama
45 men
8. Cuci seperti ad 2
9. Tambahkan substrat OPD/NPP
10. Analisis di bawah mikroskop inverted
6. ELISA (Enzyme linkage
immunosorbance assay)

Prinsip:
a. Ag + Ab (dilabel
alkaline phospatase/
peroxydase)
b. Ag + Ab + Konjugate
(dilabel Alkaline
Label
phospatase/
Ab 2 peroxydase
Ab 1
Protein

Sel ELISA, MTT, NASBA


Sel
IMMUNOFLUORSCENCE
5. Immunofluorescen
(direct and indirect IF)
Prinsip : Ag + Ab
(dilabelFITC/Rhodamin)

Ag + Ab + Konjugate
(Fab IgG) dilabel
FITC/Rhodamin

Imunohistokimia
Imunositokimia
In situ -
Hybridisation
IMMUNJOFLUORECENCE
REACTIVITY ADAPTIVE IMMUNITY
USING MARKER CD4,CD8,CD14
SIGNALING TRANSDUCTION IN
CELLULAR IMMUNE RESPONS
Goose Muskaf
Pegeon

Duck Chicken

Peripheral blood mononuclear cells (PBMCs) from animal


experiment were reacted using toll like receptors as
signaling reseptor (Fedik AR et al 2006)
AGAR GEL IMMUNODIFUSION
DESIGN AGID
AGAR GEL
IMMIUNODIF
USION (AGID)
AGID
Complemen fixation test CFT
WESTERN BLOT
1. SDS-
PAGE, 2.
WESTRN
BLOT

Anda mungkin juga menyukai