Rapid diagnosis of thalassemias and other hemoglobinopathies by capillary electrophoresis system

Winichagoon, Pranee; Svasti, Saovaros; Munkongdee, Thongperm; Chaiya, Wantana; Boonmongkol, Piatip; Chantrakul, Nawarath; Fucharoen, Suthat

doi:10.1016/j.trsl.2008.08.004

Cited by 46 publications

(32 citation statements)

References 19 publications

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“…Anemia in β-thalassemia is due to both ineffective erythropoiesis and reduced lifespan of circulating effete red blood cells, stemming from the presence of toxic unmatched α-hemoglobins, which are believed to cause apoptosis of erythroid precursor cells within the bone marrow and premature eryptosis of the circulating thalassemic red blood cells [3,4]. In fact, there are other causes of anemia, such as microenvironment factors.…”

Section: Introductionmentioning

confidence: 99%

Cytokine-Induced Apoptosis of Beta-Thalassemia/Hemoglobin E Erythroid Progenitor Cells via Nitric Oxide-Mediated Process in vitro

Kheansaard

Panichob²,

Fucharoen³

et al. 2011

Acta Haematol

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Background/Aim: β-Thalassemia/hemoglobin E (β-thal/HbE) is a common hereditary anemia in Thailand. Ineffective erythropoiesis due to apoptosis and decreased lifespan of circulating thalassemic red blood cells are the major causes of anemia. Changes to bone marrow microenvironment could contribute to apoptotic events. This study examined the effects of cytokines interleukin-1β, tumor necrosis factor-α and interferon-γ on apoptosis of β-thal/HbE erythroid progenitor cells in vitro, including nitric oxide-mediated apoptotic processes. Methods: Percent apoptosis of erythroid progenitor cells from 5 β-thal/HbE patients and 5 normal control subjects was examined using flow cytometry. In addition, the inducible nitric oxide synthase (iNOS) mRNA level and nitrite production were measured using quantitative PCR and the Griess method, respectively. Results: Upon cytokine treatment, a higher percent apoptosis was obtained with β-thal/HbE erythroid progenitor cells compared with control, and the maximum effect was observed using 20 ng/ml interferon-γ on day 14 of culture. There was an increase in iNOS mRNA level and a concomitant elevation of nitrite concentration in culture medium. Apoptosis and nitrite level were abrogated when β-thal/HbE and control cells were treated with S-methylisothiourea sulfate, an iNOS inhibitor. Conclusion: The marked sensitivity of erythroid progenitor cells from β-thal/HbE patients to cytokine-induced apoptosis via an NO-mediated process reflects a proapoptotic status of such thalassemic red blood cells.

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Section: Introductionmentioning

confidence: 99%

Cytokine-Induced Apoptosis of Beta-Thalassemia/Hemoglobin E Erythroid Progenitor Cells via Nitric Oxide-Mediated Process in vitro

Kheansaard

Panichob²,

Fucharoen³

et al. 2011

Acta Haematol

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“…-thalassemia. This phenomenon was also observed using the capillary electrophoresis system [6]. Although the developed ELISA could be used to classify b-thalassemia heterozygotes as well as the reference HPLC method, the percentages of HbA 2 determined by the developed ELISA were lower than those obtained from the HPLC method in all subject groups.…”

Section: Discussionmentioning

confidence: 64%

“…Currently, HbA 2 level is generally accepted as a principle diagnostic feature for screening of b-thalassemia heterozygotes [2]. At present, the relative quantity of HbA 2 in RBCs can be estimated by several laboratory methodologies, including electrophoresis, anion-exchange chromatography, high performance liquid chromatography (HPLC), low pressure liquid chromatography (LPLC) and capillary electrophoresis (CE) [5][6][7][8][9][10][11]. Currently, the anion-exchange microchromatographic and HPLC techniques are accepted as the reference methods for quantifying HbA 2 levels [2,5,7,12].…”

Section: Introductionmentioning

confidence: 99%

Sandwich ELISA for hemoglobin A2 quantification and identification of β-thalassemia carriers

et al. 2010

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Hemoglobin (Hb) A2 (alpha2delta2) is a minor hemoglobin in human red blood cells. An abnormal increase in the level of HbA2 is the most significant parameter in the diagnosis of beta-thalassemia carriers. In this study, we produced two monoclonal antibodies (mAbs) that specifically react to the delta-globin chain of HbA2. A sandwich type ELISA was developed employing the produced anti-HbA2 mAbs. HbA2 levels quantified by the developed sandwich ELISA were highly correlated with those obtained from the standard HPLC method (r = 0.934, p < 0.001). HbA2 levels determined by the ELISA were 4.4 +/- 1.9% in beta-thalassemia heterozygotes compared to 1.4 +/- 0.8, 1.9 +/- 0.8, 1.5 +/- 0.8 and 1.5 +/- 0.6% in normal subjects, HbE heterozygotes, suspected alpha-thalassemia heterozygotes and HbE homozygotes, respectively. Using a cut-off value of 2.5%, beta-thalassemia heterozygotes could be separated from non-beta-thalassemia heterozygotes with the same accuracy as obtained using the standard HPLC method. More importantly, the developed ELISA was able to determine HbA2 levels in HbE-bearing individuals which could not be done by the HPLC method. Our results suggest that this sandwich ELISA can be applied for mass screening for beta-thalassemia heterozygotes, especially in resource-limited countries, where beta-thalassemia is highly prevalent.

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“…La variación de hemoglobina A2 en electroforesis en geles de agarosa se ha descrito en estudios como el de Clarke y Higgins, donde comparan EC y HPLC y concluyen que este tipo de electroforesis carece de exactitud y precisión al detectar hemoglobina A2 22,23 . La EC ofrece ventajas operativas frente a la electroforesis en gel de agarosa: mínima cantidad de muestra, realización de diluciones precisas, no requiere lavado de eritrocitos, rapidez y automatización.…”

Section: Discussionunclassified

Variantes de hemoglobina en una población con impresión diagnóstica positiva para hemoglobinopatías en Colombia

et al. 2015

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Rapid diagnosis of thalassemias and other hemoglobinopathies by capillary electrophoresis system

Cited by 46 publications

References 19 publications

Cytokine-Induced Apoptosis of Beta-Thalassemia/Hemoglobin E Erythroid Progenitor Cells via Nitric Oxide-Mediated Process in vitro

Cytokine-Induced Apoptosis of Beta-Thalassemia/Hemoglobin E Erythroid Progenitor Cells via Nitric Oxide-Mediated Process in vitro

Sandwich ELISA for hemoglobin A2 quantification and identification of β-thalassemia carriers

Variantes de hemoglobina en una población con impresión diagnóstica positiva para hemoglobinopatías en Colombia

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