JENIS-JENIS METODE

IDENTIFIKASI VIRUS
PADA BAHAN
PEMERIKSAAN KLINIS
RAPID TEST

ELISA

PCR

SPEKTROFOTOMETRI
 ELISA
(Enzyme Immunoassay and Enzyme-
Linked Immunosorbent Assay)
 Prinsip ELISA
Reaksi dasar imunologi:
antigen berikatan dengan antibodi spesifik
(Antigen dalam jumlah sedikit dapat dideteksi)
Proses:
antigen (protein, hormon, peptide) dalam bentuk fluida/cairan diikat oleh
antibodi spesifik dilabeli lagi oleh antibodi kedua (enzyme coupled- antibody)
perubahan warna/fluoresensi (dari perubahan substrat kromogenik) yang
mengindikasikan adanya antigen (Gbr. 1)
Pengamatan:
perubahan warna (intensitas dan gradasi warna yang timbul) menunjukkan adanya
antigen. Semakin tua/kuat warna yang timbul maka jumlah antigen semakin tinggi
yyang dikukur absorbansinya
 Pendahuluan
 Alat diagnostik kualitatif dan kuantitatif dalam bidang obat dan industri

 Radioimmunoassay (RIA) : was first described by Berson and Yalow (Yalow

and Berson, 1960)

kekurangan: radioisotop yang berbahaya

 Enzyme immunoassay (EIA) and enzyme-linked immunosorbent assay (ELISA)

keunggulan: menggunakan enzim (lebih aman dibandingkan dengan
radioisotop)
 Fungsi ELISA
 ELISA is a biochemical assay that uses antibodies and an enzyme-mediated
color change to detect the presence of either antigen (proteins, peptides,
hormones, etc.) or antibody in a given sample.

 The competitive method detects compositional differences in complex antigen

mixtures with high sensitivity, even when the specific detecting
antibody is present in relatively small amounts.

 Multiple and portable ELISA is a ready-to-use, low-cost lab kit that is ideal for
large population screening in low-resource settings.
Cara Kerja ELISA
 Jenis-jenis ELISA
1.Direct ELISA

2.Indirect ELISA

3.Sandwich ELISA

4.Competitive ELISA

5.Multiple and portable ELISA

 Indirect ELISA advantages

 High sensitivity: More than one labeled antibody

is bound per antigen molecule;

 Flexible: Different primary detection antibodies

can be used with a single labeled secondary
antibody;

 Cost-saving: Fewer labeled antibodies are

required.
 Disadvantages of direct ELISA

The primary antibody must be labeled individually,

which can be time-consuming and inflexible when
performing multiple experiments.

Also, the signal is less amplified in direct ELISA,

which means a lower sensitivity and could be viewed
as a disadvantage to some.
 Sandwich ELISA advantages:

High specificity, since two antibodies are used the antigen/analyte is

specifically captured and detected

Suitable for complex samples, since the antigen does not require purification
prior to measurement

Flexibility and sensitivity, since both direct and indirect detection methods can
be used
 Competitive ELISA advantages:
 High specificity, since two antibodies are used the
antigen/analyte is specifically captured and detected
 Suitable for complex samples, since the antigen does
not require purification prior to measurement
 Flexibility and sensitivity, since both direct and indirect
detection methods can be used