4

Sistem Penggolongan Darah

Efrida, dr., SpPK., MKes

4 Desember 2014
 Tujuan Pembelajaran
Mampu menjelaskan:
1.Dasar pembagian golongan darah
(terutama sistem ABO dan Rhesus)
2.Prinsip pemeriksaan golongan darah
ABO dan Rhesus
3.Kepentingan menentukan golongan
darah
 Dasar Pembagian Golongan
Darah
Konsep dasar Imunohematologi:
1.Seroimunologi, genetik, biokimia
2.Studi molekular antigen terkait dengan
struktur membran pada komponen selular
darah
3.sifat-sifat serta reaksi imunologi seluruh
komponen darah dan konstituennya
Susunan Darah
Volume darah dewasa: 46 liter
Terdiri atas plasma dan sel-sel darah (eritrosit,
leukosit, trombosit)

Eritrosit/the red blood cells (RBCs):

mengandung hemoglobin, suatu protein yang
mengikat oksigen. RBCs mentranspor oksigen ke
jaringan dan mengangkut karbondioksida dari
jaringan.

Leukosit/the white blood cells: respons imun

selular terhadap infeksi.

Trombosit/the platelets: peran pada

hemostasis primer (membentuk sumbat
trombosit pada luka pembuluh darah)

Plasma: mengandung berbagai substansi

terlarut (protein, lipid, elektrolit)
Eritrosit
Struktur
Bikonkaf, anukleat
Komponen
Hemoglobin
Lipid, ATP, karbonik
anhidrase
Fungsi
Transpor oksigen
dari paru ke jaringan
dan karbondioksida
dari jaringan ke
paru-paru

19-5
 ABO antigen Biochemistry

Carbohydrate
ABH antigens
expressed on
RBC
glycoproteins
&
glycosphingoli
pid (type 2,3,4
chain) RBC
origin.

Type 1 chain are synthesized by gastrointestinal mucosa secreted into plasma

passively adsorbed onto RBC membrane
Glikoprotein dan glikolipid pada
permukaan eritrosit:
- substansi selular bersifat spesifik
- diatur secara genetik (Mendelian)
- bersifat imunogenik
- mampu merangsang pembentukan
aloantibodi spesifik bila masuk ke
dalam tubuh seseorang yg tidak
memiliki substansi tsb
 Blood Grouping

Determined by antigens
(agglutinogens) on surface of RBCs
Antibodies (agglutinins) can bind to
RBC antigens, resulting in
agglutination (clumping) or
hemolysis (rupture) of RBCs

19-10
 Erythrocyte Antigens
& Antibodies

> 700 antigens organized

into 29 blood group systems
by the International Society of
Blood Transfusion (ISBT).
 Sistem penggolongan terpenting:
1. ABO
2. Rhesus
Sistem lain diperiksa jika antigennya
menimbulkan reaksi transfusi yg tidak
disebabkan gol ABO atau Rhesus, misal:
- Sistem Kell
- Sistem Duffy dan Kidd
- Sistem MN, P, Lewis, Lutheran (reaksi
transfusi jarang/ringan)
 Dasar Pembagian
Golongan Darah Sistem ABO
1901: Landsteiner menemukan 2 jenis
antigen pada permukaan eritrosit
- antigen (aglutinogen) A
- antigen (aglutinogen) B
Pengamatannya berdasarkan:
Reaksi aglutinasi eritrosit dan adanya
antibodi (aglutinin) pada serum
seseorang yang bereaksi langsung
terhadap antigen eritrosit
 ABO Antigens
Also express in many tissues, body
fluids, platelet and endothel
Most important blood group system in
transfusion and organ transplantation.
3 antigens: A, B, H (biosynthetic
precursor of A & B antigens).
A & B: autosomal codominant antigens
expressed on group A, B & AB RBC
 ABO Antibodies

Weak or absent in newborn 3-6 mo

5-10 yo adult level
Advancing age slight decrease
Detected at room temperature, with optimal
reactivity at 40C.
Merupakan suatu IgM atau IgG atau IgA
IgG (reactive at 370C) can occur after
transfusion/pregnancy; higher titer
Can fix complement hemolysis in vivo/vitro
Can cause: hemolytic transfusion reaction &
hemolytic disease of the new born.
Less common ABO
antibodies

Anti-A1
Anti-H
 Cont.
Berdasarkan antigen A,B pada permukaan
eritrosit dan antibodi ABO pada plasma
Sistem ABO (golongan darah ABO)

Von Decastello dan Sturli menemukan

golongan darah ke-4: AB

Penemuan selanjutnya: 2 jenis antigen A

(A1 dan A2)6 fenotipe golongan darah
 Cont
Golongan darah ABO
1. A1 (genotipe A1/A1, A1/A2, A1/O)
2. A2 (genotipe A2/A2, A2/O)
3. A1B (genotipe A1/B)
4. A2B (genotipe A2/B)
5. B (genotipe B/B, B/O)
6. O (genotipe O/O)
AB0 blood grouping system

Blood group A
If you belong to the blood
group A, you have A antigens
on the surface of your RBCs
and B antibodies in your
blood plasma.
Blood group B
If you belong to the blood
group B, you have B antigens
on the surface of your RBCs
and A antibodies in your
blood plasma.
Blood group AB
If you belong to the blood group
AB, you have both A and B antigens
on the surface of your RBCs and no
A or B antibodies at all in your
blood plasma.
Blood group O
If you belong to the blood group O (null),
you have neither A or B antigens on the
surface of your RBCs but you have both A
and B antibodies in your blood plasma.
 ABO inheritance and genetics

The ABO gene is autosomal (the gene is not on either

sex chromosomes)

The ABO gene locus is located on the chromosome 9.

A and B blood groups are dominant over the O blood group

A and B group genes are co-dominant

Each person has two copies of genes coding for their ABO
blood group (one maternal and one paternal in origin)
AUTOSOMAL CHROMOSOME

The alleles for Blood

group are in the same
A place on the B
chromosome 9.
However the genes
have a different code
giving the different father
blood group
mothe
r

one alleles from father and one

from mother
Possible Blood group Genotypes

Parent A B O
Allele
A

O
Possible Blood group Genotypes

Parent A B O
Allele
A AA AB AO

B AB BB BO

O AO BO OO
The ABO blood groups

The table shows the four ABO phenotypes ("blood groups")

present in the human population and the genotypes that give
rise to them.

Blood
Antigens
Grou Antibodies in Serum Genotypes
on RBCs
p

A A Anti-B AA or AO
B B Anti-A BB or BO
AB A and B Neither AB
O Neither Anti-A and anti-B OO
Golongan darah sistem
Rhesus (Rh)
 Cont
Golongan darah sistem Rhesus:
1.Rh + (DD, Dd)
2.Rh (dd)
 Golongan Darah Rhesus
Ciri-ciri antigennya mirip dengan sistem ABO
Antibodi imun (tidak alamiah), terbentuk kr
reaksi imunologik (transfusi atau
kehamilan/transfusi fetomaternal)
Dapat menyebabkan reaksi ketidakcocokan
pada transfusi atau kehamilan
Fisher dan Race: sistem rhesuscampuran
antigen yg kompleks, diatur oleh very closely
linked genes pada kromosom 1
 Cont
3 kelompok gen: C, D, E
Tiap kelompok bersamaan dg beberapa
alel
- C dg c dan cw
- D dg d dan Du
- E dg e
Ag D: terkuat (sangat imunogenik)
Ag Du: subgrup lemah Ag D (deteksi dg
antiglobulin tdk langsung, setelah
eritrosit diinkubasi dengan anti-D)
The Rh(D) Antigen
RH is the most complex
system, with over 45 antigens
Discovered in 1940 after work
on Rhesus monkeys
Subsequently discovered to be
unrelated to monkeys
RH gene located on short arm
of chromosome 1

ABO & Rh(D) 32

 Simple Genetics of Rh(D)
86% of caucasians are Rh(D) pos
The antithetical antigen d has not
been found
The d gene is recessive:
Dd, dD, DD, persons are Rh(D) pos
Only dd persons are Rh(D) neg

ABO & Rh(D) 33

Distribution of Rh(D) Types

Population Rh(D) pos Rh(D) neg

Caucasian 86% 14%

African- 95% 5%
American
Oriental >99% <1%

ABO & Rh(D) 34

 Significance of Rh(D)
80% of Rh(D) neg persons exposed to
Rh(D) pos blood will develop anti-D
Anti-D can also be stimulated by
pregnancy with an Rh(D) positive baby
Sensitisation can be prevented by the use of
anti-D immunoglobulin, antenatally and post
natally
Rh(D) neg females of childbearing
potential should never be given Rh(D)
positive blood products
ABO & Rh(D) 35
 Inheritance
ABO & RH genes are not linked
ABO & Rh(D) type are inherited
independently
For example:
An A Rh(D) positive mother
and a B Rh(D) positive father
could have an O Rh(D) negative child

ABO & Rh(D) 36

 Inheritance of ABO and
Rh(D)
Mother Father

Group A AO Group B BO
Rh(D) pos Dd Rh(D) pos Dd

Group A AO Group B BO Group O OO

Rh(D) pos Dd Rh(D) pos Dd Rh(D) neg dd

ABO & Rh(D) 37

Rh Blood Group and Rh Incompatibility
A person with Rh- blood does not have Rh antibodies
naturally in the blood plasma

Blood Alleles
Genotype
Type Produced

DD D
Rh positive
Dd D or d

Rh negative dd d
Do you know which blood group you
belong to?

According to above blood

grouping systems, you can
belong to either of
following 8 blood groups:
A person with Rh- blood can develop Rh
antibodies in the blood plasma if he or she
receives blood from a person with Rh+ blood,
whose Rh antigens can trigger the production
of Rh antibodies.

A person with Rh+ blood can receive blood

from a person with Rh- blood without any
problems.
 Aplikasi Klinis
Sistem Penggolongan Darah
 History of Blood Groups and Blood Transfusions

Experiments with blood transfusions

have been carried out for hundreds of
years. Many patients have died and it was
not until 1901, when the Austrian Karl
Landsteiner discovered human blood
groups, that blood transfusions became
safer.

He found that mixing blood from two

individuals can lead to blood clumping.
The clumped RBCs can crack and cause
toxic reactions. This can be fatal.
http://nobelprize.org/medicine/educational/landsteiner/readmore.html
History of Blood Groups and Blood
Transfusions (Cont.)

Karl Landsteiner discovered that blood

clumping was an immunological reaction
which occurs when the receiver of a blood
transfusion has antibodies against the donor
blood cells.

Karl Landsteiner's work made it possible to

determine blood types and thus paved the
way for blood transfusions to be carried out
safely. For this discovery he was awarded the
Nobel Prize in Physiology or Medicine in
1930.
 Kepentingan Klinis
Transfusi darah dan transplantasi organ
Identifikasi dan pencegahan terhadap
aloimunisasi ibu hamil oleh antigen Rhesus
Menentukan diagnosis, prognosis, dan
terapi HDN (hemolytic disease of the
newborn) akibat aloantibodi
Diagnosis dan pemeriksaan destruksi
eritrosit yg disebabkan autoantibodi atau
aloantibodi
 Kedokteran kehakiman
antropologi
 Prinsip Pemeriksaan
Golongan Darah Sistem
ABO dan Rhesus
 Prinsip: Imunoserologi
(Reaksi Antigen-Antibodi)
Membran Eritrosit: protein dan karbohidrat

Antigen Eritrosit

Antibodi spesifik
Penampilan Golongan Darah
ABO
Uji golongan darah harus dilakukan
terhadap :
Eritrosit
Serum
Pada keadaan meragukan harus
diulang
Several methods for testing the ABO group of an
individual exist. The most common method is:

Serology: This is a direct detection of the ABO

antigens. It is the main method used in blood
transfusion centres and hospital blood banks.

This form of testing involves two components:

a) Antibodies that are specific at detecting a

particular ABO antigen on RBCs (forward
method)

b) Cells that are of a known ABO group that

are agglutinated by the naturally occurring
antibodies in the person's serum (reverse
 Pemeriksaan Golongan Darah
ABO
1. Forward Grouping
- Menentukan gol darah ABO
berdasarkan
antigen pada eritrosit
- Aglutinasi antigen bereaksi
dengan
antisera (IgM anti-A atau anti-B).
Anti-A antigen A
Anti-B antigen B
When RBCs carrying one or both antigens are exposed to the
corresponding antibodies, they agglutinate; that is, clump together.
People usually have antibodies against those red cell antigens that
they lack.

Human RBC before (left) and after (right) adding serum

containing anti-A antibodies. The agglutination reaction
reveals the presence of the A antigen on the surface of
the cells.
 Anti AB dan Anti A
Anti-AB dianjurkan diuji pada
penggolongan darah donor
Anti-A mempunyai gabungan 2
antibodi
Anti-A yang mengaglutinasi sel-sel A1,
A2, A1B dan A2B
Anti-A1 yang hanya mengaglutinasi sel-
sel A1 dan A1B
Prosedur kerja:
Dibuat suspensi eri 2-4% dg NaCl 0.9%
Disediakan 3 tabung & diberi label: A, B,
Tabung A di(+) 1 tetes antiserum anti-A
Tabung B di(+) 1 tetes antiserum anti-B
Tabung AB di(+) 1 tetes antiserum anti-A
Masing2X tabung di(+) 1 tetes suspensi e
Ketiga tabung disentrifus 15 dg 3400 rp
Resuspensi & dilihat ada aglutinasi/ tidak
Interpretasi hasil:
Antisera Interpretasi Gol Darah
Anti-A Anti-B Anti-AB
0 0 0 O
+ 0 + A
0 + + B
+ + + AB
0 0 + Subgolongan A lemah

Keterangan:
+ = Aglutinasi
0 = Tidak ada aglutinasi
 2. Reverse Grouping
- Menentukan gol darah ABO
berdasarkan
ada/tidak adanya antibodi
- Aglutinasi antibodi dalam
serum
bereaksi dengan antigen A/B
pd eritrosit
Prosedur kerja:
Dua buah tabung diberi label: A, dan B
Masing2x tabung di(+) 2 tetes serum/
plasma
yg akan diperiksa
Ke dalam tabung A di(+) 1 tetes reagen e
A1
Ke dalam tabung B di(+) 1 tetes reagen e
B
Kedua tabung disentrifus 15 dg 3400 rp
Resuspensi & dilihat ada aglutinasi/ tidak
Interpretasi hasil:

A1 B Antibodi Golongan Darah

+ + Anti-A & Anti-B O

0 + Anti-B A
+ 0 Anti-A B
0 0 Tidak ada AB

Keterangan:
+ = Aglutinasi
0 = Tidak ada aglutinasi
 Illustration of the forward and reverse
grouping reaction patterns of the ABO
groups using a blood group tile

http://www.bh.rmit.edu.au/mls/subjects/abo/resources/genetics1.htm
 Metode Pemeriksaan
Metode Slide
Prosedur kerja:
Pemeriksaan forward grouping:
Pada kaca objek ditaruh 1 tetes reagen anti-
A,
anti-B, & anti-AB
Darah diteteskan pada reagen itu & dicampu
Kaca objek digoyang perlahan-lahan
Dilihat apakah ada aglutinasi
Pemeriksaan reverse grouping:
Pada kaca objek ditaruh 1 tts reagen eri
A1&B
Serum/ plasma diteteskan pd reagen itu
& dicampur dengan ujung lidi
Kaca objek digoyang perlahan-lahan
Dilihat apakah ada aglutinasi
Interpretasi Hasil:
Forward Grouping
Antisera Golongan Darah
Anti-A Anti-B Anti-AB
0 0 0 O
+ 0 + A
0 + + B
+ + + AB

Keterangan:
+ = Aglutinasi
0 = Tidak ada aglutinasi
 Reverse Grouping

A B Golongan Darah

+ + O
0 + A
+ 0 B
0 0 AB

Keterangan:
+ = Aglutinasi
0 = Tidak ada aglutinasi
 Keuntungan:
Cepat, mudah, sederhana, memerlukan
sedikit sampel.

Kerugian:
1) tidak dpt menentukan aglutinasi lemah

2) tidak ada kontrol (+), kontrol (-), &

autokontrol
 Metode Tube
Pemeriksaan eri & serum scr
bersamaan.
Pemeriksaan atas eritrosit:
Dipakai 12 tbg kecil, msg2x diisi 1 tts NaCl
0.9%
Di(+) 1 tetes susp 2% eri X, eri A, eri B, & eri
O
pada masing-masing baris
Di(+) 1 tetes anti-A, anti-B, & anti-AB pd
masing
masing kolom, dicampur
Hasil dibaca setelah 2 jam
Pemeriksaan atas serum:

Dipakai 4 tabung reaksi kecil

Tiap tabung diisi 1 tetes NaCl 0.9%
Tabung ke-1 ditambah suspensi 2% eri A
Tabung ke-2 ditambah suspensi 2% eri B
Tabung ke-3 ditambah suspensi 2% eri X
Tabung ke-4 ditambah suspensi 2% eri O
Eritrosit & zat anti dicampur
Hasil dibaca setelah 2 jam
Interpretasi hasil:
Golongan Darah A
Atas Eri Atas
Serum

+ O + Eri X O Eri A

+ O + Eri A + Eri B

Auto
O + + Eri B O Eri X
kontrol

O O O Eri O O Eri O

Anti A Anti B Anti AB

Interpretasi hasil:
Golongan Darah B
Atas Eri Atas
Serum

O + + Eri X + Eri A

+ O + Eri A O Eri B

Auto
O + + Eri B O Eri X
kontrol

O O O Eri O O Eri O

Anti A Anti B Anti AB

Interpretasi hasil:
Golongan Darah AB
Atas Eri Atas
Serum

+ + + Eri X O Eri A

+ O + Eri A O Eri B

Auto
O + + Eri B O Eri X
kontrol

O O O Eri O O Eri O

Anti A Anti B Anti AB

Interpretasi hasil:
Golongan Darah O
Atas Eri Atas
Serum

O O O Eri X + Eri A

+ O + Eri A + Eri B

Auto
O + + Eri B O Eri X
kontrol

O O O Eri O O Eri O

Anti A Anti B Anti AB

 Keuntungan:
1) Pemeriksaan eritrosit dan serum
dilakukan secara bersamaan
2) Ada kontrol (+), kontrol (-), dan
autokontrol
3) Dapat menentukan aglutinasi lemah
4) Dapat dilakukan pemeriksaan imun
antibodi

Kerugian:
Sukar & memerlukan waktu lama
 Metode Gel Test
Didasarkan reaksi antigen &
antibodi
Reaksi antigen & antibodi terjadi
di dalam
tabung kecil yang berisi gel
Partikel aglutinasi dipisahkan dari
partikel
non-aglutinasi oleh gel.
Prosedur kerja:

Dibuat suspensi sel (tanpa dicuci) dg ID

Diluent (Modified Low Ionic Strength
Solution),
500 L diluent:5 L darah, dicampur
Suspensi sel dipipet ke dlm tabung berisi
gel yang mengandung antibodi spesifik:
50 L
ID Card diinkubasi 37C, 15 menit
ID Card disentrifus selama 10 menit
Hasil Reaksi dibaca secara makroskopis
Interpretasi hasil:
+4 : Menggumpal & menyatu shg
cairan tampak jernih
+3 : Menggumpal tp tdk menyatu. Jd
tdd beberapa gumpalan kasar,
sekitarnya tampak cairan jernih
+2 : Gumpalan agak kasar tp tdk
semua sel darah beraglutinasi,
tampak cairan agak keruh
+1 : Gumpalan halus, tampak cairan
keruh
Keuntungan:
Sederhana, mudah
Hasil reaksi stabil
Tdk ada fase pencucian
Hanya memerlukan sedikit
sampel
Pembacaan reaksi secara
makroskopis
Masa kadaluarsa reagen
panjang
Mengurangi limbah
Meningkatkan keamanan
laboratorium
Kerugian:
Memerlukan peralatan yang mah
Memerlukan waktu >
lama dr M. slide