Sistem Penggolongan Darah dan

Aplikasi untuk Transfusi

Efrida, dr., SpPK., MKes

Maret 2014
 Tujuan Pembelajaran
Mampu menjelaskan:
1. Dasar pembagian golongan darah
berdasarkan sistem ABO dan Rhesus
2. Prinsip pemeriksaan golongan darah
ABO dan Rhesus
3. Kepentingan menentukan golongan
darah
 Dasar Pembagian
Golongan Darah
1900-an: Landsteiner menemukan 2 jenis
antigen pada permukaan eritrosit
- antigen (aglutinogen) A
- antigen (aglutinogen) B
Pengamatannya:
Reaksi aglutinasi eritrosit dan adanya
antibodi (aglutinin) pada serum seseorang
yang bereaksi langsung terhadap antigen
ini
 Cont.
Berdasarkan hal tsb 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)
 Cont
Golongan darah sistem Rhesus:
1. Rh + (DD, Dd)
2. Rh (dd)
 Basic Immunohematology
Concept

Immunohematology:
Serologic, genetic, biochemical, and
molecular study of antigens associated
with membrane structures on the
cellular constituents of the blood, and
immunologic properties & reactions, of
all blood components and constituents.
 History
Karl Landsteiner
(1900) golongan darah

26 sistem golongan darah

Paling penting: ABO
Rhesus
 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.
What is blood made up of?

An adult human has about 46 liters of blood

circulating in the body.
Blood consists of several types of cells floating
around in a fluid called plasma.

The red blood cells (RBCs) contain

haemoglobin, a protein that binds oxygen. RBCs
transport oxygen to, and remove carbon dioxide
from the tissues.

The white blood cells fight infection.

The platelets help the blood to clot, if you get

a wound for example.

The plasma contains salts and various kinds of

proteins.
Erythrocytes
Structure
Biconcave,
anucleate
Components
Hemoglobin
Lipids, ATP, carbonic
anhydrase
Function
Transport oxygen
from lungs to tissues
and carbon dioxide
from tissues to lungs
19-13
 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
Groups
ABO and Rh

19-14
 Erythrocyte Antigens &
Antibodies
> 700 antigens organized into 29
blood group systems by the
International Society of Blood
Transfusion (ISBT).
 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).
6 phenotypes
A & B: autosomal codominant antigens
expressed on group A, B & AB RBC
 ABO antigen Biochemistry
Carbohydrate
ABH antigens
expressed on
RBC
glycoproteins &
glycosphingolip
id (type 2,3,4
chain) RBC
origin.

Type 1 chain are synthesized by gastrointestinal mucosa secreted into plasma

passively adsorbed onto RBC membrane
 ABO Antibodies
Weak or absent in newborn 3-6 mo
5-10 yo adult level
Advancing age slight decrease
Detected at room temperature, saline agglutinins
with optimal reactivity at 40C.
Mostly IgM.
IgG (reactive at 370C) can occur after
transfusion/pregnancy; higher titer; less readily
neutralized by soluble blood group substances.
Can fix complement hemolysis in vivo/vitro
Can cause: hemolytic transfusion reaction &
hemolytic disease of the new born.
Perkembangan antigen-antigen A dan B sel
darah merah
Perkembangan antigen eritrosit sejak
janin s/d remaja.
Bayi baru lahir (s/d 3 bulan) : reaksi lebih
lemah dibanding dewasa, terutama
antibodi
Jadi pengujian pada BBL cukup eritrosit
saja
Less common ABO
antibodies

Anti-A1
Anti-H
What are the different blood groups?
The differences in human blood are due to the
presence or absence of certain protein
molecules called antigens and antibodies.

The antigens are located on the surface of the

RBCs and the antibodies are in the blood
plasma.

Individuals have different types and

combinations of these molecules.

The blood group you belong to depends on

what you have inherited from your parents.
ABO blood grouping system

According to the ABO blood

typing system there are four
different kinds of blood types:
A, B, AB or O (null).
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 Mustafa
blood group
Sara

one alleles from Mustafa and

one from Sara.
What do co-dominant genes mean?
This meant that if a person inherited one A group gene
and one B group gene their red cells would possess
both the A and B blood group antigens.

These alleles were termed A ( which produced the A

antigen ),
B (which produced the B antigen) and O (which was "non
functional"and produced no A or B antigen)
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 most important in assuring a safe blood transfusion.

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
Why group A blood must never be
given to a group B person?

Giving someone blood from the wrong ABO

group could be fatal.

The anti-A antibodies in group B attack group

A cells and vice versa.

Blood group O negative is a different story.

The Rhesus (Rh) System
 Golongan Darah Rhesus
Ciri-ciri antigennya mirip dengan sistem
ABO
Dapat menyebabkan reaksi ketidak
-cocokan 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) 38

 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) 39

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) 40

 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) 41

 Inheritance
ABO & RH genes are not linked
ABO & Rh(D) type are inherited
independently
For example:
An A Rh(D) pos mother
and a B Rh(D) pos father
could have an O Rh(D) neg child

ABO & Rh(D) 42

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) 43

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.
Any questions?
The ABO Blood Group System

Laboratory Determination of
the ABO System
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.

b) Cells that are of a known ABO group that

are agglutinated by the naturally occurring
antibodies in the person's serum.
 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
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.

http://users.rcn.com/jkimbal
 Penampilan Golongan Darah
ABO
Uji golongan darah harus dilakukan
terhadap :
Eritrosit
Serum
Pada keadaan meragukan harus
diulang
 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
Membran eritrosit Karbohidrat
Protein

Antigen Golongan Darah

Antibodi spesifik
Antigen eritrosit:
Stabil seumur hidup
Pada beberapa kondisi berubah

Antibodi eritrosit:
Dibentuk secara alamiah, bukan
karena pemaparan thd eritrosit
Pada beberapa kondisi berubah
Perubahan antigen & antibodi
diskrepansi ABO

Hasil golongan darah pada:

forward grouping

reverse grouping
Diskrepansi ABO harus diatasi
karena dapat berakibat fatal:
transfusi darah
transplantasi organ

Penentuan gol darah juga berperan

untuk:
kedokteran kehakiman
antropologi
Diskrepansi sistem ABO
Definisi
Perbedaan hasil golongan darah ketika
dilakukan pemeriksaan cara forward
grouping dg reverse grouping

Etiologi
Kesalahan teknis
Macam-macam kondisi klinis/penyakit
 Masalah yang dihadapi dalam
penggolongan darah secara sistem
ABO
Ekspresi antigen ABO dapat berkurang
Perubahan antigen ABO
ABO alloantibodi
Adanya aglutinin yang tidak diharapkan
Populasi eritrosit yang heterogen
Kesalahan pembacaan uji golongan darah
Polyagglutinable erythrocytes
Kesalahan teknis positif palsu adalah:

Pelabelan tabung sampel salah

Reagen yang terkontaminasi/ tabung
yang digunakan untuk pemeriksaan
kotor
Overcentrifugation
Interpretasi atau pencatatan hasil salah
Kesalahan teknis negatif palsu adalah:

Pelabelan tabung sampel salah

Sampel/ reagen yang digunakan waktu
pemeriksaan jumlahnya kurang
Undercentrifugation
Suhu inkubasi tidak tepat
Reagen yg digunakan sudah terlalu lama
Interpretasi & pencatatan hasil salah
Secara klinis, penyebab diskrepansi ABO
digolongkan menjadi 4 kategori yaitu:
Melemahnya antigen/ perubahan antigen
Reaksi antigen yang tidak diharapkan
Melemahnya antibodi atau perubahan
antibodi
Reaksi antibodi yang tidak diharapkan
 Melemahnya/ perubahan antigen
Melemahnya subgolongan antigen A/ B
Penyakit menetap: leukemia
Antigen A/ B yg terlarut jumlahnya
berlebihan dlm plasma
Transplantasi sumsum tulang dg ABO
non identik
Transfusi RBC dg ABO non identik
 Reaksi antigen yang tdk diharapkan
Antigen B didapat
Perubahan antigen
Eritrosit diselubungi antibodi
Zat aditif pada antisera (akriflavin)
Pencampuran darah
Reaksi antigen dan antibodi rendah
Melemahnya/ perubahan antibodi
Umur
Hipogamaglobulinemia
Agamaglobulinemia kongenital/ didapat
Chimerism
Transplantasi sumsum tulang dg ABO
non identik
Reaksi antibodi yg tdk diharapkan
Formasi Rouleaux
Cold autoantibodies
Isoaglutinin yang tidak diharapkan
Antibodi yang tidak diharapkan
 Pemeriksaan Golongan Darah ABO
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
Prosedur kerja:
Dibuat suspensi eri 2-4% dg NaCl 0.9%
Disediakan 3 tabung & diberi label: A, B, & AB
Tabung A di(+) 1 tetes antiserum anti-A
Tabung B di(+) 1 tetes antiserum anti-B
Tabung AB di(+) 1 tetes antiserum anti-AB
Masing2X tabung di(+) 1 tetes suspensi eri
Ketiga tabung disentrifus 15 dg 3400 rpm
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
 Reverse Grouping
Menentukan gol darah ABO
berdasarkan ada/ tidak adanya antibodi
Aglutinasi antibodi dalam serum
bereaksi dengan antigen A/ B pd eri
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 eri A1
Ke dalam tabung B di(+) 1 tetes reagen eri B
Kedua tabung disentrifus 15 dg 3400 rpm
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
 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 & dicampur
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

A1 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 mahal
Memerlukan waktu > lama dr M. slid
Sekian dan terima kasih