Capillary Zone Electrophoresis Developed for Rapid Determination of Thalassemia
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By LabMedica International staff writers Posted on 10 Jun 2015 |
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Image: The P/ACE MDQ Capillary Electrophoresis System (Photo courtesy of Beckman Coulter Inc.).
A capillary zone electrophoresis (CZE) method has been developed for human globin determination in the diagnosis of thalassemia and hemoglobin variants as analysis of globin chains is an important aspect of thalassemia diagnostics.
Thalassemia is an inherited autosomal recessive blood disorder characterized by the underproduction of globin chains as a consequence of globin gene defects, resulting in malfunctioning red blood cells and oxygen transport.
Medical scientists at the Southern Medical University (Guangzhou, China) evaluated an assay on blood samples obtained from anticoagulated whole blood from a total of 310 clinical samples consisting of 58 normal, 107 α-thalassemia silent/trait, 42 β-thalassemia carriers and 63 β-thalassemia major and β-thalassemia intermedia (β -TM/TI), and 40 others with various hemoglobinopathies.
Hemoglobin analysis was carried out using the high-performance liquid chromatography (HPLC, Variant II, Bio-Rad Laboratories; Hercules, CA, USA). Capillary zone electrophoresis (CZE) was performed using a P/ACE MDQ Capillary Electrophoresis System (Beckman Coulter Inc.; Fullerton, CA, USA) equipped with a 70 cm × 50 μm uncoated fused-silica capillary at 25 °C.
Distinct globin peaks were resolved in 17 minutes and coefficients of variation (CV) for migration time and areas ranged from 0.37% to 1.69% and 0.46% to 6.71%, respectively. Receiver operating characteristic (ROC) curve analysis of the α/β area ratios gave 100% sensitivity and specificity for indicating β-TI/TM, and 100% sensitivity and 97.4% specificity for Hb H disease. Hemoglobin G-Honolulu (Hb G-Honolulu) and Hb Westmead (Hb WS) were successfully detected using this CZE method.
The authors concluded that they had successfully developed a simple, rapid, high-resolution CZE method for the separation of globin chains using highly acidic buffer and uncoated capillaries. Using this method, α-, β- and γ-chains, and αWS and αG Honolulu chains were successfully separated within 17 minutes, indicating suitability for routine clinical applications. The migration times and globin chain peak areas were highly reproducible; although not as rapid as existing high-throughput HPLC techniques. This CZE method may provide an attractive complementary and/or confirmatory approach. The study was published in the June 2015 issue of the journal Blood Cells Molecules and Diseases.
Related Links:
Southern Medical University
Bio-Rad Laboratories
Beckman Coulter Inc.
Thalassemia is an inherited autosomal recessive blood disorder characterized by the underproduction of globin chains as a consequence of globin gene defects, resulting in malfunctioning red blood cells and oxygen transport.
Medical scientists at the Southern Medical University (Guangzhou, China) evaluated an assay on blood samples obtained from anticoagulated whole blood from a total of 310 clinical samples consisting of 58 normal, 107 α-thalassemia silent/trait, 42 β-thalassemia carriers and 63 β-thalassemia major and β-thalassemia intermedia (β -TM/TI), and 40 others with various hemoglobinopathies.
Hemoglobin analysis was carried out using the high-performance liquid chromatography (HPLC, Variant II, Bio-Rad Laboratories; Hercules, CA, USA). Capillary zone electrophoresis (CZE) was performed using a P/ACE MDQ Capillary Electrophoresis System (Beckman Coulter Inc.; Fullerton, CA, USA) equipped with a 70 cm × 50 μm uncoated fused-silica capillary at 25 °C.
Distinct globin peaks were resolved in 17 minutes and coefficients of variation (CV) for migration time and areas ranged from 0.37% to 1.69% and 0.46% to 6.71%, respectively. Receiver operating characteristic (ROC) curve analysis of the α/β area ratios gave 100% sensitivity and specificity for indicating β-TI/TM, and 100% sensitivity and 97.4% specificity for Hb H disease. Hemoglobin G-Honolulu (Hb G-Honolulu) and Hb Westmead (Hb WS) were successfully detected using this CZE method.
The authors concluded that they had successfully developed a simple, rapid, high-resolution CZE method for the separation of globin chains using highly acidic buffer and uncoated capillaries. Using this method, α-, β- and γ-chains, and αWS and αG Honolulu chains were successfully separated within 17 minutes, indicating suitability for routine clinical applications. The migration times and globin chain peak areas were highly reproducible; although not as rapid as existing high-throughput HPLC techniques. This CZE method may provide an attractive complementary and/or confirmatory approach. The study was published in the June 2015 issue of the journal Blood Cells Molecules and Diseases.
Related Links:
Southern Medical University
Bio-Rad Laboratories
Beckman Coulter Inc.
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