Khera Rachna, Singh Tejinder, Khuana Nita, Gupta Naresh, Dubey A P
Department of Pathology, Maulana Azad Medical College, New Delhi, 110002 India.
Department of Medicine, Maulana Azad Medical College, New Delhi, India.
Indian J Hematol Blood Transfus. 2015 Mar;31(1):110-5. doi: 10.1007/s12288-014-0409-x. Epub 2014 Jun 5.
High-performance liquid chromatography (HPLC) is a technique introduced for the accurate diagnosis of hemoglobinopathies and thalassemias. The advantage of the HPLC system is the excellent resolution, reproducibility & quantification of several normal & abnormal hemoglobin resulting in accurate diagnosis of thalassemia syndromes. The purpose of this study is to evaluate the HPLC technique in diagnosis of thalassemia syndromes and also correlate it with clinicohematological profile in these cases. A total of 110 cases were diagnosed as thalassemias and hemoglobinopathies by Bio- Rad variant II HPLC system by β-thal short program. The retention times, proportion of the haemoglobin (%), and peak characteristics for all hemoglobin (Hb) fractions were recorded. Alkaline Hb electrophoresis was performed in each case. Other tests performed were HbF estimation by Betke's method, brilliant cresyl blue preparation for HbH inclusion bodies, sickling tests using 2 % metabisulphite and serum Ferritin estimation. Family studies were carried out wherever necessary. Of 110 cases included in the study, 87 cases were of thalassemic disorders and 23 cases were of hemoglobinopathies. Four Hb variants were identified including HbD, HbE, HbS, HbJ Oxford. There was a significant decrease in the level of HbA2 associated with iron deficiency anemia. The mean HbA2 levels in both iron deplete and iron replete groups were clearly >4 %, suggesting that HPLC identified nearly all high HbA2 β-thalassemia trait even in spite of iron deficiency.
高效液相色谱法（HPLC）是一种用于准确诊断血红蛋白病和地中海贫血的技术。HPLC系统的优点是对几种正常和异常血红蛋白具有出色的分辨率、重现性和定量能力，从而能够准确诊断地中海贫血综合征。本研究的目的是评估HPLC技术在诊断地中海贫血综合征中的应用，并将其与这些病例的临床血液学特征相关联。通过Bio-Rad Variant II HPLC系统的β-地中海贫血短程序，共诊断出110例地中海贫血和血红蛋白病。记录所有血红蛋白（Hb）组分的保留时间、血红蛋白比例（%）和峰特征。对每个病例进行碱性Hb电泳。进行的其他检测包括用贝特克法估算HbF、用煌焦油蓝制备HbH包涵体、用2%偏亚硫酸氢钠进行镰变试验以及血清铁蛋白估算。必要时进行家系研究。在纳入研究的110例病例中，87例为地中海贫血症，23例为血红蛋白病。鉴定出四种Hb变异体，包括HbD、HbE、HbS、HbJ Oxford。与缺铁性贫血相关的HbA2水平显著降低。缺铁组和铁充足组的平均HbA2水平均明显>4%，这表明即使存在缺铁，HPLC仍能识别出几乎所有高HbA2β-地中海贫血特征。
