2

Thalassemia screening test

..
. .
2553
(thalassemia) (abnormal Hb Hb variant)
globin chain
Hb Hb autosomal
recessive 2 alleles
allele (thalassemia trait, carrier, heterozygote)
500,000
10,000

Hb
Hb 98% O2
4 subunits subunit heme globin
chain quaternary structure
heme porphyrin ring Fe Fe O2
heme synthesis porphyria
globin chain 2
1. like subunit 141 -globin chain () -globin chain ()
gene 16
2. -like subunit 146 -globin chain (), -globin chain (), -globin
chain () -globin chain () gene 11
Hb Hb A (22) ~97%, Hb A2 (22) ~2.5% Hb F (22) <1%

Hb

Hb 2
1. globin chain ()
thalassemia -globin -thalassemia -globin
-thalassemia
-thalassemia
-globin genotype /
-thalassemia deletion 2 genes -thalassmia 1 (--/)
-globin -thalassemia 2 (- /)
-globin

-thalassemia

-thalassemia -thalassemia point mutation

-globin gene -globin 0-thalassemia
+-thalassemia
globin chain globin chain
globin chain
(ineffective erythropoiesis)

(chronic hemolysis)
2. abnormal Hb point mutation

- Hb E missense mutation -globin gene codon 26 GAG AAG
glutamate lysine cryptic splice site
codon 25 normal splice site codon 30 cryptic splice site premature stop
condon -globin RNA normal splice site -globin
26 -globin Hb E Hb
-globin
- Hb Constant Spring (Hb CS) mutation -globin gene (CS) codon 142
TAA stop codon CAA glutamine translation
stop codon CS-globin 141 172 amino
acid Hb Constant Spring Hb

Hb
Hb barts hydrops fetalis genotype homozygous thal 1 (--/--) -globin Hb F (22)
-globin Hb Barts (4) oxygen affinity

(hydrops fetalis)

Hb H disease genotype -thal 1/ -thal 2 (--/- ) -thal 1/Hb CS (--/ CS )

-globin -globin -chain 4
Hb H Hb H inclusion bodies
reticuloendothelial system Hb Barts

Hb H inclusion bodies
(hemolytic crisis)
2

Homozygous -thalassemia genotype 0/0, +/+

-globin 0 + -globin
Hb Hb F (22) Hb A
(22) -globin Hb A2
Hb F
thalassemic facies
20 ,

-thalassemia/ Hb E disease genotype 0/E +/E

-globin
Homozygous -thalassemia

(pigment
stone)
-thal 1 trait (--/), -thal 2 trait (-/), Homozygous -thal 2 (-/-), Hb CS trait(CS/
), -thalassemia trait (/0, /+), Hb E trait (/ E) Homozygous Hb E (E/E)

Screening test

3
1. MCV (mean corpuscular volume)
80 fL
( MCV )
2. one tube osmotic fragility test (OFT) hypotonic saline
solution (0.36% NaCl)


3. Hb DCIP (dichlorophenol-indophenol precipitation
test) DCIP Hb Hb
Hb E Hb H, Hb Barts
2 MCV OFT DCIP

1. Hb typing or Hb analysis electrophoresis high performance liquid chromatography

1
2. DNA analysis PCR -thalassemia 1 -thalassemia 2

3. serum ferritin,
serum iron total iron binding capacity (TIBC)

5
folic acid
(stem cell transplantation)

stem cell

0/0 thalassemia
2-4 Hb

hypersplenism ()

Hb Barts hydrops fetalis

Hb H disease
Hb H Constant Spring
Homozygous Hb CS
-thal 1 trait
-thal 2 trait

Homozygous -thalassemia
-thalassemia/Hb E

Homozygous Hb E
Hb E trait

Genotype
(--/--)

Hb typing
Cord blood Hb Barts >80%
Cord blood Hb Barts >20%
(--/-)
>1 Hb H =4-20%
CS
(--/ )
Hb CS =2-3%, Hb H = 10-15%
CS
CS
( / )
Hb CS~5%
Cord blood Hb Barts =2-10%
(--/)
>1 normal Hb typing
Cord blood Hb Barts <4%
(-/)
Normal Hb typing
0 0
+ +
( / ), ( / ) Hb A2 Hb F
(0/+)
Absent or low level of Hb A
0 E
( / )
Hb F Hb A Hb E
( + / E )
(E/E)
Hb E =80-100%
E
( / )
Hb E =25-35%, Hb A

(/0)
Hb A , Hb A2>3.5%
(/+)
1 genotype Hb typing Hb
0/+ compound heterozygote Homozygous
-thalassemia
-thal trait

MCV DCIP test

(. Clinical practice guidelines for diagnosis and management of

thalassemia syndrome, 2006.)


1. , , , . .
: , 2544
2. . 2006. Clinical practice guidelines for diagnosis and
management of thalassemia syndrome. (cited 24 Nov 2009). Available from:
http://www.thalassemia.or.th/thal-cpg.pdf.
3. , , , . 1999. KKU-DCIP-Clear reagent
kit for HbE screening. (cited 12 Feb 2010). Available from:
http://www.iat.or.th/inventor/invention_doc/100013842.pdf.
4. Prayongratana K, Polprasert C, Raungrongmorakot K, Tatone K, Santiwatanakul S. Low cost
combination of DCIP and MCV was better than that of DCIP and OF in the screening for Hemoglobin
E. J Med Assoc Thai 2008;91:1499-1504.