Advanced Course in Blood Transfusion

12-13 April Usi Sukorini, Dep Patologi Klinik & Ked. Lab FK UGM/ Instalasi
2017 Laboratorium Klinik RSUP Dr. Sardjito, Yogyakarta
Suatu glikoprotein yang tersusun atas 4
rangkaian polipeptida yang saling berikatan
melalui ikatan disulfida
Antibody
• glycoprotein (immunoglobulin) that recognizes
a particular epitope on an antigen and
facilitates clearance of that antigen

Immunoglobulin
• Antibody; glycoprotein secreted by plasma
cells that binds to specific epitopes on
antigenic substances
Jenis Antibodies Tipe Ig IgG, IgM, IgA, IgD, IgE

hangat
Suhu reaktivitas
dingin

complete
Kemampuan
mengglutinasi eritrosit
incomplete

reguler
klasifikasi ABO & non
ABO
ireguler
 Goldar: Deteksi &
1 • ABO
• Rh
2 Identifikasi
Antibodi
3 Uji silang
serasi
Sistim ABO  tidak terlibat dlm pem.
skrining antibodi
 Antigen make up

 Sistim ABO
 Sistim Rhesus
 Non ABO

Skrining Ab  non ABO !

Doses Effect

Sistem non ABO

Doses Effect

Sistem non ABO

 ANTIBODY DETECTION

• Antibody Screen
• Autocontrol
• Direct Antiglobulin Test
• Potentiators
• Patient history
 Antibody screen

 Indikasi: deteksi Ab terhadap RBC

 Misalnya pada:
 Pasien membutuhkan transfusi
 Wanita hamil atau sesudah melahirkan
 Pasien dg suspek reaksi transfusi
 Donor darah dan plasma
 Konsep skrining antibodi

Menginkubasi
serum/
plasma dg
sel skrining
IAT IgG
pd 37⁰C

+ sel skrining
 SKRINING ANTIBODI

SERUM PASIEN
(tidak diketahui ada/tdk & jenis
Ab-nya)
SEL SKRINING
(diketahui susunan antigennya)
 Antibody Detection and Identification

Purpose:
• The purpose of the antibody screen is to detect red blood
cell antibodies other than anti-A or anti-B.
• These antibodies are called “unexpected” or irreguler Ab
because only 0.3 to 2 % of the general population have
positive antibody screen.
• Once an unexpected antibody is detected, antibody
identification studies are performed to determine the
antibodies specificity and clinical significance.
 Antibody Screening

PRINCIPLE:
1. Detect unexpected clinically significance
antibodies
2. Ab screen test consist of:
Testing the recipient serum or plasma against two
or three screening cells of known antigen
composistion
3. Red cell antibodies may cause direct
agglutination or lysis of red cells, or may coat
the red cell with globulin (eg. IgG, C3)
 Antibody Screening

PRINCIPLE:
4. Screening cells are incubated with recipient
serum or plasma at 37°C
5. After incubation the cells are observed for
direct agglutination or hemolysis
6. Washed to removed unbound globulin and
tested with antihuman globulin serum (AHG)
 Antibody Screening

PRINCIPLE:

7. Direct agglutination or hemolysis usually indicates

the presence of IgM (cold antibodies)
8. Agglutination with AHG indicates that the screening
cells have been coated with IgG and/or C3)
9. Some of the clinically significance antibodies usually
detected by AHG phase are:
 Anti- Rh
 Anti-Kell
 Anti-Duffy
 Anti-Kidd
 MNS system (anti-S, anti-s)
When identification Ab is determine?

 Antibody Screens use 2 or 3 Screening Cells

to “detect” if antibodies are present in the
serum

 If antibodies are detected, they must be

identified…
 Antibody screening test involve testing patient’s
serum against two or three reagent red blood
cell samples called screening cells

 Screening cells are commercially prepared group

O cell suspensions obtained from individual
donors that are phenotype for the most
commonly encountered and clinically important
red blood cell antigens.
 Antibody Screening

• Group O cells are used so that naturally occurring

anti-A or anti-B will not interfere with detection of
unexpected antibodies.

• The cells are selected so that the following antigens

are present minimally:
D, C, E, c, e, M, N, S, s, P, Lua, Lub , K, k, Lea, Leb, Fya,
Fyb, Jka and Jkb
ANTIBODY SCREENING: Ab DETECTION

 KNOWN • UNKNOWN
 Screening cells • Serum
 ANTIBODY IDENTIFICATION

KNOWN  UNKNOWN
 Panel cells  Serum
 Reagent RBCs

 Screening Cells and Panel Cells are the same with

minor differences:
 Screening cells (panel skrining)
 Antibody detection
 Sets of 2 or 3 vials

 Panel cells (panel identifikasi)

 Antibody identification
 At least 10 vials per set
 Skrining Antibodi (deteksi antibodi)

 Panel kecil
 Autocontrol (AC)

 AC menentukan jenis Ab (alloantibodi atau

autoantibodi)
 AC = suspensi eritrosit pasien + serum pasien
 Jika AC + dan DAT (-):
 Potentiator  menyebabkan positif palsu
 Hrs diulang dg potentiator berbeda, atau tanpa
mengggunakan enhancement solution
 AC + / DAT + : autoAb
 Warm or cold type
 Phases

• Fase atau temperatur reaksi  aglutinasi tampak sbg

indikasi bhw antibodi tsb adalah IgG atau IgM
• IgM: bereaksi pd temperatur kamar atau IS
• Misalnya: anti-Lea, anti-Leb, anti M, anti N anti-I dan
anti P1 perlu dicurigai jika reaksi IS terdeteksi

• IgG: bereaksi pd fase antiglobulin

• Reaksi pd fase berbeda  > 1 Ab dan kombinasi
antibodi IgG dan IgM
 Kekuatan reaksi

• Dlm panel, semua reaksi kuat dan dlm kekuatan yang

mirip
• Antibodi: anti-K, anti-D, anti-E, anti anti-e, anti-c dan
anti-C bersifat > kuat dp anti-Fy, anti-Fyb, anti-Jka,
anti-Jkb, anti-S dan anti-s
• Kekuatan reaksi juga bervariasi dg ‘dosis’ antigen
• Jika panel homosigot  reaksi lebh kuat
 Ruling out (ekslusi)

• Pengertian: mengeksklusi antigen yg diekspresikan oleh sel

(pd sel panel), jika hasil tes negatif (0) atau dengan kata
lain sel panel yg memberi hasil (antara serum + sel panel)
reaksi negatif (0) dapat diekslusi

• Urutan melakukan ruling out:

1. Dipilih hasil reaksi negatif pertama antara serum dgn sel panel
2. Hasil negatif tsb di atas  dicocokan/diplotkan dengan
spesifisitas Ag yg + pd sel panel (tdp antigen yg diekspresikan
oleh sel panel).
3. Jika reaksi Ag-Ab tidak terjadi (negatif=0) berarti Ab tidak
bereaksi dg sel panel, shg Ab dpt dieliminasi sbg Ab yg terkait
 Ruling out (ekslusi)

4. Sel panel heterosigot terutama Duffy, Kidd, dan

sistem MNS, tidak dieksklusi krn Ab terlalu lemah
utk bereaksi
5. Selanjutnya ruling out (eksklusi) sel panel yang
memberikan reaksi negatif kedua dst
6. Proses eksklusi akan mengerucutkan kemungkinan
antibodi yang ada
 Matching the pattern

• Selanjutnya dilihat reaksi positif dan di-match-kan

dgn polanya
• Jika terdapat single antibody, maka pola reaksi yg
dihasilkan di-match-kan dgn kolom antigen
• Misalnya aglutinasi tjd dengan sel 1,3 dan 7; dan
antigen K tdp pd sel maka Ab diidentifikasikan sbg
anti K
 Rule of three

 Ditujukan utk membuat kesimpulan bhw hasil bukan

merupakan suatu kebetulan
 Harus tdp minimal 3 hasil positif dan 3 hasil negatif
 Jika belum mencapai ‘rule of three’  sel panel
ditambah
 Secara statistil: Fischer exact test
 The Fischer exact test

 To ensure that the paterrn or reactivity does not

result from chance alone, whenever possible the
panel should contain 3 cells positive for antigen in
question and 3 cells negative for the antigen.
ANTIBODY IDENTIFICATION
Langkah analisis dan interpretasi
1. Ruling out (eksklusi)
2. Circle antigens not crossed out
3. Consider antibody’s usual reactivity
3. Matching the pattern
 4. Rule of three (at least 3 positive & 3 negative)

Apabila ‘rule of three’’ blm tercapai  tambah sel panel

Analisis: - didapatkan antibodi ireguler anti-K yang bereaksi pada suhu 37 C,

- anti Lua belum dapat disingkirkan
TERIMA KASIH