Clinical Biochemistry 44 (2011) 406–411

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Clinical Biochemistry
j o u r n a l h o m e p a g e : w w w. e l s e v i e r. c o m / l o c a t e / c l i n b i o c h e m

Thalassemia and hemoglobinopathies in Southeast Asian newborns: diagnostic

assessment using capillary electrophoresis system
Hataichanok Srivorakun a,b, Goonnapa Fucharoen b, Yossombat Changtrakul c,
Patcharee Komwilaisak d, Supan Fucharoen b,⁎
a
Biomedical Science Program, Graduate School, Khon Kaen University, Khon Kaen, Thailand
b
Centre for Research and Development of Medical Diagnostic Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University, Khon Kaen, Thailand
c
Clinical Microscopy Unit, Srinagarind Hospital, Khon Kaen University, Khon Kaen, Thailand
d
Department of Pediatrics, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand

a r t i c l e i n f o a b s t r a c t

Article history: Background: We have investigated the Capillarys 2 Hemoglobin testing system to assist in presumptive
Received 4 November 2010 diagnosis of thalassemia and hemoglobinopathies commonly found in Southeast Asia.
Received in revised form 3 January 2011 Methods: Study was conducted on 226 newborns. Hematological parameters were recorded and Hb
Accepted 14 January 2011 profiles were examined on the Capillarys 2 Hemoglobin analyzer (SEBIA). DNA analyses were used to
Available online 28 January 2011
establish the final diagnoses.
Results: Among 226 newborns examined, 122 had thalassemias with 17 different genotypes. The capillary
Keywords:
Newborns screening
electrophoresis system could provide useful data for presumptive diagnoses of cases, especially those with Hb
Thalassemia E and α-thalassemia. Hb E was found to be 2.6–6.2% in heterozygote whereas Hb Bart's were clearly observed
Hemoglobinopathy in cases with compound heterozygous or homozygous α+-thalassemia and heterozygous α0-thalassemia. Hb
Capillary electrophoresis H disease and other forms of α-thalassemia could be differentiated based on the presence of Hb Bart's and its
percentage.
Conclusion: The capillary electrophoresis system is applicable to newborn screening for common forms of
thalassemia in Southeast Asia.
© 2011 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Introduction severe thalassemia. Diagnosis at newborn is useful for early treatment

Thalassemia and hemoglobinopathies are inherited hemoglobin
(Hb) disorders found worldwide. In Thailand and other Southeast Asian
countries, the disease is very common with 20–30% of the population
having the α-thalassemia trait, 3–9% the β-thalassemia trait and 20–30%
the Hb E trait. Heterozygous forms of these genetic defects are
asymptomatic with very mild hypochromic microcytosis. However,
with such a carrier rate, complex interactions between them result in a
wide spectrum of clinical syndromes of α- and β-thalassemias
commonly encountered in the regions [1,2]. Diagnosis of the diseases
can be performed at various stages of development i.e. prenatal,
neonatal, and adult periods. Identification of thalassemia in the adult
stage is crucial for treatment and prevention and control of the disease.
Prenatal diagnosis is an important step in prevention of new cases with
⁎ Corresponding author at: Centre for Research and Development of Medical
Diagnostic, Laboratories, Faculty of Associated Medical Sciences, Khon Kaen University,
Khon Kaen 40002, Thailand. Tel./fax: +66 43 202 083.
E-mail address: supan@kku.ac.th (S. Fucharoen).
planning and prevention in the next child. The methods usually applied
include alkaline electrophoresis, isoelectric focusing (IEF) and high
performance liquid chromatography (HPLC) [3,4]. The Capillarys 2 Hb
testing system (Sebia) is a relatively new automated Hb analyzer which
uses the principle of capillary zone electrophoresis (CZE) [5]. With this
technique, charged Hb molecules are separated by their electrophoretic
mobilities in alkaline buffer with a specific pH. Separation also occurs
according to the electrolyte pH and electro osmotic flow created from
negative charges on the capillary wall that shorten the analysis duration
[6]. Several studies have evaluated this system and have shown
excellent precision and accurate quantification of normal Hbs and
identification of major Hb variants such as Hbs S, C and E [7–9]. We have
also demonstrated that fetal Hb analysis using this capillary electro-
phoresis system can be an effective alternative to DNA assay for prenatal
diagnosis of severe thalassemia diseases in Southeast Asia [10]. In this
study, we investigate the ability of this capillary electrophoresis system
to identify and make presumptive diagnosis of various forms of
thalassemia commonly encountered among Southeast Asian newborns.
Hematological features of these thalassemic newborns, including a
hitherto un-described condition of double heterozygote for Hb E and Hb
Constant Spring/Hb Paksé are also presented.

0009-9120/$ – see front matter © 2011 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
doi:10.1016/j.clinbiochem.2011.01.006
 H. Srivorakun et al. / Clinical Biochemistry 44 (2011) 406–411
Fig. 1. Representative capillary electrophoregrams of newborns with normal genotype (A), heterozygous α+-thalassemia (B), heterozygous Hb Constant Spring (C), homozygous α+-thalassemia (D), heterozygous α0-thalassemia (E) and Hb
H disease (F).

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408 H. Srivorakun et al. / Clinical Biochemistry 44 (2011) 406–411

Materials and methods
Subjects and hematological analysis
Ethical approval of the study protocol was obtained from the
Institution Review Board of Khon Kaen University, Thailand (HE
510728). Left-over blood specimens anti-coagulated with EDTA were
obtained from 226 newborns, aged 0–7 days, encountered at the
Diagnostic Microscopy Unit, Srinagarind Hospital, and the thalassemia
service unit of our center at Khon Kaen University for routine
hematological examination. Erythrocyte parameters were recorded on
the Coulter-STKS automated blood cell counter (Coulter Electronics,
Hialeah, Fla., USA). Hb fractions and quantifications were performed using
automated capillary zone electrophoresis (Capillarys 2: Sebia, Lisses,
France) [10]. Hematological values were presented as mean±standard
deviation or as raw data where appropriate.
DNA analyses
Genomic DNA was prepared from peripheral blood leukocytes of the
patients using the standard method. Identification of the α0-thalassemia
(SEA and THAI deletions), α+ - thalassemia (3.7 and 4.2 kb deletions),
Hb Constant Spring and Hb Paksé were routinely performed in our
laboratory using PCR methods described elsewhere [11,12]. The βE and
β-thalassemia mutations were identified using allele specific PCR assays
[13,14].
Results
Of the 226 newborns examined, 122 (53.9%) were found to carry
thalassemia genes. No thalassemia gene was detected in the remaining
104 cases (46.1%). As many as 17 different genotypes were observed.
This result confirms the high prevalence and heterogeneity of
thalassemia and hemoglobinopathies in the region. Hb analysis of
these 104 presumably normal newborns using capillary electrophoresis
revealed two major peaks of Hb F and Hb A. A small but notable peak of
Hb A2 with the amount of 0.2 ± 0.2% could be observed in 55 newborns
(Fig. 1A) but not in the remaining 49 cases (Table 1). Twenty newborns
were found to be heterozygous α+-thalassemia with 3.7 kb deletion.
Minute amounts of Hb Bart's (0.4–1.0%) (Fig. 1B) were observed in 5 of
them. In contrast, among 16 newborns with Hb Constant Spring
(Fig. 1C), this Hb Bart's was identified in 14 cases although Hb Constant
Spring (~ 0.2%) was observed in only 3 of them. Higher amounts of Hb
Bart's (3.3–6.2%) were clearly observed in all cases with compound
heterozygous for α+-thalassemia and Hb Constant Spring, homozygous
α+-thalassemia and heterozygous α0-thalassemia (Fig. 1D and 1E). As
shown in Fig. 1F, further increases of Hb Bart's levels were observed
together with small peaks of Hb H (0.7 and 0.9%) in newborns with Hb H
disease (19.0%) and Hb H-Constant Spring disease (28.6%) (Table 1).
Again, Hb Constant Spring was not detected in the latter cases, accurate
diagnosis can only be obtained after DNA analysis.
Fig. 2 demonstrates the electrophoregrams of Hb analysis for
newborns with various Hb E related disorders. As the Capillarys 2 system
can report Hb E in the presence of Hb A2, accurate levels of both Hbs could
be obtained. As shown in Table 2, in 51 pure heterozygotes, the level of Hb
E was found to be in the range of 2.6–6.2% whereas small amounts of Hb A2
(0.2±0.1%) may or may not be observed. Similar patterns were recorded
for those of double heterozygotes for Hb E / α+-thalassemia with either
3.7 kb deletion (−α3.7) or Hb Constant Spring (αCSα) (Fig. 2A–C). Hb
Bart's was only detected in some cases. A lower production of Hb E with
increased Hb Bart's level (~5.0%) was observed in newborns with Hb E /
double heterozygous α+-thalassemia or Hb E / α0-thalassemia (Fig. 2D
and E). Interestingly, further reduction of Hb E (1.2 and 3.0%) with higher
proportions of Hb Bart's (12.4 and 15.4%) with the presence of small
amounts of Hb H (0.3 and 0.4%) were observed for newborns with Hb E/
homozygous Hb Constant Spring or Hb E/compound heterozygous Hb
Constant Spring and Hb Paksé, a hitherto undescribed condition (Fig. 2F).
As shown in Table 2, although hematologically these conditions could not
be differentiated from others, the results indicate a more imbalanced
globin chain synthesis for these unusual cases and that these gene
interactions could lead to the Hb H disease in newborns.
Fig. 3A demonstrates the electrophoregram of newborns with
homozygous Hb E in which only Hb E (8.3–13.2%) and Hb F (91.7 and
91.6%) but not Hb A were observed. As for heterozygous Hb E,
interaction of this homozygous Hb E with Hb Constant Spring was
associated with a lower proportion of Hb E (6.7%) and a small amount
of Hb Bart's (0.5%), while no peak of Hb Constant Spring was found
(Fig. 3B).
Our results from 226 newborns demonstrate that the most prevalent
genotype in these Southeast Asian newborns is heterozygous Hb E (51
cases), followed by heterozygous α+-thalassemia (20 cases), heterozy-
gous Hb Constant Spring (16 cases), double heterozygous Hb E /α+-
thalassemia (9 cases), double heterozygous Hb E /Hb Constant Spring (7
cases), heterozygous α0-thalassemia (3 cases), double heterozygous Hb E/

Table 1
Hemoglobin profiles, hematologic parameters and thalassemia genotypes of 148 newborns with and without α-thalassemia.

Genotype (no.) Hb type (no.) Hb fraction (%) Hematologic parameters

Hb A2 Hb F Hb A Hb Bart's Rbc (× 1012/L) Hb (g/dL) Hct (%) MCV (fl) MCH (pg) RDW (%)
A A
αα/αα, β /β (104) A2FA (55) 0.2 ± 0.2 76.0 ± 6.8 23.7 ± 6.5 – 4.9 ± 0.7 17.4 ± 2.7 48.4 ± 7.2 97.6 ± 4.0 35 ± 1.2 17.2 ± 1.7
FA (49) – 85.7 ± 5.5 14.3 ± 5.4 – 4.9 ± 0.7 17.2 ± 2.3 48.2 ± 6.0 99.8 ± 8.7 35.6 ± 2.7 17.2 ± 1.7
-α3.7/αα, βA/βA (20) A2FABart's (1) 0.3 60.3 38.4 1.0 6.1 19.0 54.6 89.8 31.3 18.1
FABart's (4) – 80.1 ± 4.2 19.4 ± 4.3 0.4 ± 0.05 5.2 ± 0.8 17.0 ± 2.8 48.3 ± 7.9 92.5 ± 3.4 32.6 ± 1.5 17.4 ± 2.1
A2FA (6) 0.2 ± 0.2 75.1 ± 6.3 24.7 ± 6.2 – 4.8 ± 0.6 15.2 ± 2.1 44.3 ± 5.6 92.0 ± 3.9 31.5 ± 1.3 18.6 ± 2.7
FA (9) – 84.8 ± 5.0 15.2 ± 5.0 – 5.2 ± 0.4 16.9 ± 1.3 47.9 ± 2.9 92.4 ± 5.7 32.5 ± 1.4 17.6 ± 2.2
αCS α/αα, βA/βA (16) CSA2FABart's (2) 0.2, 0.1 68.6, 79.8 29.7, 18.7 1.3, 1.2 5.9, 5.2 19.1, 17.5 55.1, 50.1 92.9, 95.6 32.2, 33.4 17.6, 16.1
A2FABart's (8) 0.2 ± 0.1 75.5 ± 8.2 23.0 ± 7.7 1.1 ± 0.4 5.0 ± 0.8 16.2 ± 1.9 46.4 ± 4.7 94.0 ± 7.1 32.8 ± 1.9 18.1 ± 1.7
FABart's (4) – 82.2 ± 1.3 16.4 ± 1.0 1.4 ± 0.3 4.8 ± 0.4 15.8 ± 1.1 45.7 ± 4.5 95.1 ± 7.4 32.9 ± 1.7 17.5 ± 2.0
CSA2FA (1) 0.5 58.0 41.3 – 5.2 17.9 50.4 96.9 34.4 15.8
FA (1) – 84.7 15.3 – 5.8 17.8 54.0 93.8 30.9 18.3
αCS α/-α3.7, βA/βA (2) A2FABart's (2) 0.2, 0.2 63.9, 68.7 30.8, 25 5.1, 6.2 5.4, 5.9 15.3, 15.7 48.2, 48.6 89.1, 82.7 28.3, 26.7 21.8, 20.9
-α3.7/-α3.7, βA/βA (1) A2FABart's (1) 0.1 70.0 26.6 3.3 NA NA NA NA NA NA
–SEA/αα, βA/βA (3) FABart's (3) – 79.7 ± 4.5 15.1 ± 3.5 5.2 ± 1.2 4.7 ± 0.4 13.0 ± 1.6 40.0 ± 4.9 85.4 ± 7.6 27.9 ± 2.3 19.1 ± 3.3
–SEA/-α3.7, βA/βA (1) A2FABart'sH (1) 0.1 54.7 24.1 19.0 5.60 13.8 42.1 75.2 24.6 22.4
–SEA/αCSα, βA/βA (1) CSA2FABart'sH (1) 0.1 51.7 18.1 28.6 NA NA NA NA NA NA

Values are presented as mean ± SD or as raw data where appropriate. NA; not available.
 H. Srivorakun et al. / Clinical Biochemistry 44 (2011) 406–411
Fig. 2. Representative capillary electrophoregrams of newborns with Hb E including a pure heterozygous Hb E (A), double heterozygous Hb E / α+-thalassemia (B), double heterozygous Hb E/Hb Constant Spring (C), heterozygous HbE with
homozygous α+-thalassemia (D), double heterozygous Hb E/α0-thalassemia (E) and heterozygous Hb E with compound Hb Constant Spring/Hb Paksé (F).

409
410 H. Srivorakun et al. / Clinical Biochemistry 44 (2011) 406–411

Table 2
Hemoglobin profiles, hematologic parameters and thalassemia genotypes of 78 newborns with various Hb E disorders.

Genotype (no.) Hb type (no.) Hb fraction (%) Hematologic parameters

Hb A2 Hb E Hb F Hb A Hb Bart's Rbc Hb Hct MCV MCH RDW

(×1012/L) (g/dL) (%) (fl) (pg) (%)

αα/αα, βE/βA (51) A2EFA (18) 0.2 ± 0.1 5.1 ± 2.3 82.3 ± 5.3 12.5 ± 4.0 – 4.7 ± 0.8 16.4 ± 2.9 45.6 ± 6.9 96.4 ± 4.2 34.5 ± 1.6 18.0 ± 2.6
EFA (33) – 3.0 ± 1.1 88.0 ± 3.8 9.0 ± 3.4 – 4.8 ± 0.6 17.5 ± 1.8 48.6 ± 4.6 101.2 ± 7.4 36.5 ± 2.8 17.6 ± 1.5
3.7 E A
-α /αα, β /β (9) A2EFA (5) 0.3 ± 0.2 6.8 ± 2.3 75.6 ± 5.5 17.3 ± 4.5 – 4.8 ± 0.7 15.2 ± 2.3 42.9 ± 5.9 90.2 ± 5.9 31.9 ± 1.9 16.4 ± 1.2
EFA (4) – 3.0 ± 0.8 89.7 ± 5.5 7.3 ± 4.9 – 5.4 ± 0.45 17.8 ± 1.3 49.8 ± 4.0 92.7 ± 7.2 33.1 ± 2.6 17.6 ± 1.6
αCSα/αα, βE/βA (7) A2EFABart's (5) 0.3 ± 0.3 5.3 ± 1.9 81.0 ± 5.8 12.7 ± 3.6 0.6 ± 0.3 4.6 ± 0.6 15.1 ± 2.8 43.5 ± 8.8 94.2 ± 7.8 32.7 ± 2.1 18.8 ± 4.9
EFABart's (1) – 3.5 87.2 8.4 0.9 5.0 16.6 45.9 91.6 33.1 16.4
EFA (1) – 3.0 87.3 9.7 – 5.7 18.2 50.4 88.6 32.0 19.2
αCSα/-α3.7, βE/βA (2) EFABart's (1) – 2.8 82.8 8.2 6.2 6.6 20.3 57.9 87.5 30.7 21.1
CSA2EFABart's (1) 0.1 3.3 78.3 11.3 6.6 4.8 15.0 44.5 92.3 31.1 20.8
αCSα/αPSα, βE/βA (1) CSEFABart'sH (1) – 1.2 75.3 7.0 15.4 5.9 17.8 57.4 97.1 30.1 22.2
CS CS E A
α α/α α, β /β (1) CSA2EFABart's (1) 0.1 3.0 74.5 9.7 12.4 5.1 15.8 48 94.7 31.2 23.3
–SEA/αα, βE/βA (3) EFABart's (2) – 3.0, 2.6 85.3, 83.7 7.1, 12.1 4.6, 1.6 7.1, 6.6 19.6, 19.3 57.6, 52.6 80.6, 79 27.4, 29 22.4, 22.8
A2EFABart's (1) 0.3 4.8 75.5 17.5 1.9 5.1 13.5 37.2 72.8 26.5 16.1
αα/αα, βE/βE (3) A2EF (1) 0.3 13.2 85.7 – – 3.8 13.2 38.3 100.3 34.6 21.7
EF (2) – 8.3, 8.4 91.7, 91.6 – – 5.0, 4.5 17.7, 14.9 49.9, 41.6 99.4, 92.4 35.3, 33.1 18.4, 16.9
CS E E
α α/αα, β /β (1) EFBart's (1) – 6.7 92.8 – 0.5 4.6 14.7 41.1 89.7 32.1 15.7

Values are presented as mean ± SD or as raw data where appropriate. NA; not available.

α0-thalassemia (3 cases) and homozygous Hb E (3 cases). Other
remaining interactions including an undescribed one were found in 1 or
2 cases.
Discussion
From 226 newborns examined, we observed as many as 17
genotypes, the data confirming a high prevalence and a diverse
heterogeneity of this genetic disorder in the region. Most of the cases
were α-thalassemia and Hb E related disorders. No β-thalassemia was
detected in this study. However, as mentioned above, this could be
underrepresented as β-thalassemia carriers can not generally be
diagnosed before 1 year of age because of the same Hb analysis pattern
as that of a normal infant. It has been proposed that a cutoff less than 15%
Hb A could be used for pre-selection of possible cases of β-thalassemia
for further confirmation by parent analysis [15]. We have applied this to
our group of subjects with non-α-thalassemia (Table 1, 29 with Hb A2FA
type and 17 with Hb FA type) and examined β-thalassemia mutations by
DNA analysis. No β-thalassemia mutation was detected. Further
screening for β-thalassemia was also carried out on subjects with
MCV less than 100 fl and HbA2 higher than 0.2%. No β-thalassemia
mutation was identified either (data not shown). This result confirms
the low prevalence of β-thalassemia in the region.
Unlike β-thalassemia, hematologic comparison between the non
α-thalassemic group with various forms of α-thalassemia revealed
decreasing trends for Hb, MCV, and MCH, which corresponded to the
number of α-globin gene defects (Table 2). Values were markedly
reduced and clearly observed in newborns with homozygous α+-
thalassemia, heterozygous α0-thalassemia and Hb H-disease. As for our
previous study using HPLC analysis [16], Hb Bart's could be observed in all
of them. These data indicate that identification and quantification of Hb
Bart's as well as the reduced Hb, MCV and MCH values could be used as
markers for presumptive diagnosis of these forms of α-thalassemia
genotypes. However, as for HPLC analysis, a lower level of Hb Bart's was
detected and could be missing in many cases of α+-thalassemia
heterozygotes. We observed a higher percentage of Hb Bart's for
newborns with heterozygous Hb Constant Spring as compared to
heterozygous α+-thalassemia with 3.7 kb deletion. This indicates a likely
greater globin chain imbalance for Hb Constant Spring as compared to the
deletion α+-thalassemia. Nonetheless, our data confirmed that deletion of
one α-globin gene may not be associated with a detectable amount of Hb
Bart's at birth and its identification either with electrophoresis, HPLC, IEF
or CE for diagnosis of α+-thalassemia carrier is unreliable.
As expected, the most common hemoglobinopathy found is Hb E and
its related disorders which were detected in 78 newborns (Table 2). As
shown in Figs. 2 and 3, the capillary electrophoresis system can clearly
demonstrate Hb E in all cases although Hb A2 was missing in some of
them. The lowest recognized level of Hb E was 1.2% found with the
hitherto undescribed condition of heterozygous Hb E and compound
heterozygous for Hb Constant Spring and Hb Paksé (αCSα/αPSα, βE/βA)
(Fig. 2F). All of these newborns with Hb E were positive for βE gene
screening using allele specific PCR assay [13]. This result indicates an
excellent accuracy of this Hb analysis system for identification of Hb E in
newborns. We have previously demonstrated the successful application
of this capillary electrophoresis system for identifying small amounts of
Hb E in fetal blood specimens at prenatal diagnosis [10]. Considering the
hematological data presented in Table 2 for newborns with these Hb E
related disorders, we observed a lower proportion of Hb E in accordance
with the number of α-globin genes defected i.e. the level was highest in
pure heterozygous Hb E and decreased in double heterozygous for Hb E
and α-thalassemia although with some overlapping. The level of Hb A2
did not help in diagnosis either. Again the useful marker for identifying
newborns with double heterozygous Hb E/α-thalassemia especially α0-
thalassemia and non deletional α+-thalassemia (Hb Constant Spring
and Hb Paksé) is Hb Bart's which was found in most cases with these
genotypes. However, Hb Bart's was undetectable in all newborns with
Hb E/α+-thalassemia (−α3.7/αα, βE/βA), which indicates the possibility
of mis-diagnosis of such cases on the capillary electrophoresis system.
The same findings have been noted on the HPLC and IEF formats [16,17].
It is noteworthy that in an undescribed condition of heterozygous Hb E /
Hb Constant Spring / Hb Paksé (αCSα/αPSα, βE/βA), we noted a relatively
lower level of Hb E (1.2%) and much elevation of Hb Bart's (15.4%) as
well as small peaks of Hb H and Hbs Constant Spring and Paksé on the
electrophoregram (Fig. 2F). This level of Hb Bart's, quite similar to that
detected in deletional Hb H disease (−−SEA/−α3.7, βA/βA) (Table 1;
19.0%), is much higher than those observed for the three cases of double
heterozygous Hb E and α0-thalassemia (−−SEA/αα, βE/βA) (Table 2;
1.6–4.6%). It is conceivable therefore that this rare interaction between
Hb E / Hb Constant Spring / Hb Paksé could lead to the Hb H disease in
newborns and a syndrome known as AEBart's disease commonly
encountered in thalassemia intermedia patients in northeast Thailand
[18]. As no β-thalassemia was detected in this study and it is speculated
as for HPLC and other Hb analysis methods that screening of
β-thalassemia in newborn by Hb analysis is difficult. However, analysis
using both Hb and DNA analyses, being done in this study, could provide
useful information for early diagnostic of α-thalassemia and Hb E
 H. Srivorakun et al. / Clinical Biochemistry 44 (2011) 406–411
Fig. 3. Capillary electrophoregrams of newborns with homozygous Hb E (A) and homozygous Hb E with heterozygous Hb Constant Spring (B).
related disorders, appropriate counseling to the parents and treatment
planning for the case.
In conclusion, our results demonstrate that the capillary electropho-
resis system specifically developed for Hb fraction separation and
quantification could give high resolution performance comparable to
conventional HPLC when used in newborn screening for thalassemia and
hemoglobinopathies commonly found among Southeast Asian popula-
tions. As compared to DNA assay, this Hb analysis is simple, rapid, fully
automated and would be applicable in most routine laboratories in the
regions.
Acknowledgments
The study was supported by grants to S.F. from Khon Kaen University
and the Office of Higher Education Commission (CHE-RES-RG-51),
Ministry of Education, Thailand. H.S. is supported by the Royal Golden
Jubilee PhD program (PHD/0132/2549) of the Thailand Research Fund,
Thailand. We thank Meditop Co., Ltd, Bangkok, Thailand for supplying the
Sebia Capillarys 2 electrophoresis system and reagents used in this study.
We also thank Ian Thomas for helpful comments on the manuscript.
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