© Borgis - Postępy Nauk Medycznych 2/2016, s. 88-91
Katarzyna Koza1, Adrianna Łoniewska-Lwowska1, *Jadwiga Fabijańska-Mitek1, Teresa Jackowska2, Marta Sobczyńska2, Anna Adamowicz-Salach3, Beata Burzyńska4
Diagnostyka molekularna talasemii α w polskiej populacji
Molecular diagnostics of thalassemia α in Polish population**
1Department of Immunohaematology, Centre of Postgraduate Medical Education, Warsaw
Head of Department: Jadwiga Fabijańska-Mitek, PhD, Associate Professor
2Department of Paediatrics, Centre of Postgraduate Medical Education, Warsaw
Head of Department: Teresa Jackowska, MD, PhD, Associate Professor
3Department of Paediatric Haematology and Oncology, Medical University of Warsaw
Head of Department: Prof. Michał Matysiak, MD, PhD
4Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw
Head of Institute: Prof. Teresa Żołądek, PhD
Streszczenie
Wstęp. Talasemia α jest wrodzoną niedokrwistością hemolityczną, w której mutacje w jednym lub większej ilości genów α globiny prowadzą do ograniczenia lub braku syntezy białek łańcuchów α globin. Występuje często w rejonach tropikalnych i subtropikalnych, a także jest obecnie opisywana w Ameryce Północnej i w północnej Europie. Diagnostyka molekularna pozwala niezawodnie wykrywać przypadki talasemii α. Jedną z metod jest technika ASO (wykorzystująca allelospecyficzne sondy oligonukleotydowe), wariant metody odwrotny dot-blot.
Cel pracy. Zidentyfikowanie przypadków talasemii α wśród pacjentów pediatrycznych z niezdiagnozowaną niedokrwistością hemolityczną.
Materiał i metody. Pacjenci pediatryczni z niezdiagnozowaną niedokrwistością hemolityczną rozpoznaną na podstawie parametrów klinicznych, morfologicznych (mikrocytozy) i biochemicznych badani metodą ASO.
Wyniki. 15% badanych osób okazało się przypadkami talasemii α, z najczęstszą mutacją -α3.7. Znaleziono również jedną mutację --FIL, która występuje bardzo rzadko poza Azją Południowo-Wschodnią.
Wnioski. Uzyskane wyniki wskazują, że talasemia α może być przyczyną wrodzonej niedokrwistości hemolitycznej w populacjach, w których ta choroba uznawana jest za niezwykle rzadką.
Summary
Introduction. Thalassemia α is a hereditary haemolytic anaemia, in which mutation in one or more of α globin genes leads to decrease in protein synthesis or complete absence of α globin chains. It is very common in tropical and subtropical populations, however it is now more often described in North America and northern Europe. Molecular diagnostics allows to detect cases of thalassemia α in reliable manner. One of the methods is reverse dot blot or ASO (allele-specific oligonucleotide probes) technique.
Aim. Identification of α thalassemia cases among pediatric patients with undiagnosed haemolytic anaemia.
Material and methods. Pediatric patients with undiagnosed haemolytic anaemia based on clinical, morphological (microcitosis) and biochemical parameters, tested with the use of ASO technique.
Results. Approximately 15% of the patients were α thalassemia cases, with the -α3.7 mutation being the most common. We have also found one --FIL mutation, very rare in other than Southeast Asia populations.
Conclusions. This results indicate that thalassemia α may be a cause of hereditary haemolytic anaemia in populations where this disease was considered extremely rare.
Introduction
Thalassemias are large group of haemolytic anaemias connected with mutations in one or more globin genes. This leads to decrease in protein synthesis or complete absence of globin chains. The most common are alpha (α) and beta (β) thalassemias with carrier frequency > 1% in all tropical and subtropical populations (1). Alpha thalassemia is the most frequent in Southeast Africa, the Mediterranean area, the Middle East and Africa (2). In some areas the incidence of this disorder is as high as > 40% (Middle Eastern and Indian populations) and > 80% (Papua New Guinea and small populations in north-east India) (2). However, due to accelerated migration and increased percentage of mixed marriages, thalassemias are more often described in different areas, like North America and northern Europe (1, 3-5). Improvement in molecular diagnostics allowed not only to diagnose such cases of the disorder, but also to find mutations in globin genes in indigenous European and American populations.
In human, there are four α globin genes, two on both chromosome 16. Alpha thalassemia mutations remove one of the genes on chromosome (thalassemia α+), or both of them (thalassemia α0) (2). Absence of one or two globin genes is usually asymptomatic. Lack of three of them leads to HbH disease, with anaemia of varying degrees. Absence of all α globin genes is connected with haemoglobin Bart’s hydrop fetalis, the lethal form of α thalassemia, if the mutations are α0 type (1, 5).
In Poland, thalassemias were long considered as insignificant in diagnosis of haemolytic anaemias. However, recent studies revealed presence of α and β thalassemia cases in our population (6-8).
Aim
Identification of α thalassemia cases among pediatric patients with undiagnosed haemolytic anaemia.
Material and methods
We have examined 48 pediatric patients from two pediatric departments with symptoms of haemolytic anaemia based on clinical, morphological and biochemical parameters. All patients have signed the informed consent forms, and the studies were approved by appropriate bioethical committee. The laboratory analyses included measurement of haemoglobin level (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), red blood cell distribution width (RDW), serum iron (Fe) and serum ferritin concentration.
Blood was collected using EDTA as anticoagulant and DNA was isolated with the use of QIAamp DNA Blood Mini Kit (Qiagen, Netherland). The reverse dot blot or ASO (allele-specific oligonucleotide probes) technique was used (Alpha-Globin StripAssay®, ViennaLab, Austria) to analyze mutations in α globin genes. This method allows to identify 21 of the most common mutations underlying α thalassemia.
Results
Forty eight pediatric patients with symptoms of haemolytic anaemia were examined in order to identify α thalassemia cases. The average and the range of biochemical and morphological parameters indicating presence of haemolytic anaemia in examined children are shown in table 1. Thirty one of them were male, and 17 female. The age of the patients ranged from 1.5 to 17 years.
Table 1. The average of morphological and biochemical parameters in children with microcitosis suspected of talassemia α
| Parameter normal values | Hb
11.0-14.6 g/dl | MCV
76-90 fl | MCH
25.5-32 pg | RDW
11.5-14.5% | Fe
50-175 μg/dl | Ferritin 7-140 ng/ml | HbF
1-2% | HbA2
2.5% |
| Average patients’ values | 11.52 | 68.23 | 23.13 | 15.11 | 77.9 | 46.61 | 2.26 | 2.96 |
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