Ou C N, Rognerud C L
Department of Pathology, Texas Children's Hospital and Baylor College of Medicine, 6621 Fanin Street, 77030, Houston, TX, USA.
Clin Chim Acta. 2001 Nov;313(1-2):187-94. doi: 10.1016/s0009-8981(01)00672-6.
Alkaline cellulose acetate and acidic citrate agar electrophoreses are the most widely utilized methods for hemoglobin analysis. However, due to their limited resolution, incorrect or unresolved diagnosis of common hemoglobinopathies are sometimes encountered.
Isoelectric focusing provides excellent resolution but is labor intensive and lacks accurate quantitation. High-performance liquid chromatographic methods have been developed for either screening or confirmation of hemoglobinopathies with relatively high sensitivity or specificity. Through the years, we have developed, refined and optimized an HPLC procedure using a porous silica coated with polyaspartic acid to improve the elution time of hemoglobin analysis while maintaining the high sensitivity and resolution necessary for both screening and confirmatory purposes.
The method is capable of separating more than 45 commonly encountered hemoglobin variants within 12 min. These include Barts, H, A1C, Raleigh, Hope, I, F, Camden, N-Baltimore, I-High Wycombe, I-Paris, J-Baltimore, N-Seattle, Grade, Fannin-Lubbock, Malmo, South Florida, A, Chicago, G-Georgia, Lepore-Baltimore, P-Galveston, G-Coushatta, Lepore-Boston, E, Zurich, Osu Christiansborg, A2, G-Philadelphia, Korle Bu, Russ, E-Saskatoon, Richmond, D-Punjab, Deer Lodge, Koln, Montgomery, S, Q-Thailand, G-San Jose, A2', Hasharon, Q-India, Tampa, Constant Spring, SG-hybrid, C-Harlem, O-Arab, British Columbia, and C. The method provides not only the identification of the aforementioned hemoglobin and variants but also an accurate quantitation of their concentrations, particularly Hb F and A2, which are useful for the diagnosis of HPFH and beta-thalassemia, respectively.
The simplicity of the sample preparation, superior resolution of the method, and accurate quantitation of hemoglobin concentration, combined with complete automation, make this an ideal methodology for the routine diagnosis of hemoglobin disorders in a clinical laboratory.
碱性醋酸纤维素和酸性枸橼酸盐琼脂电泳是血红蛋白分析中应用最广泛的方法。然而，由于其分辨率有限，有时会遇到常见血红蛋白病的诊断错误或无法确诊的情况。
等电聚焦提供了出色的分辨率，但劳动强度大且缺乏准确的定量分析。已经开发出了高效液相色谱方法用于血红蛋白病的筛查或确认，具有相对较高的灵敏度或特异性。多年来，我们开发、改进并优化了一种高效液相色谱程序，使用涂有聚天冬氨酸的多孔硅胶来改善血红蛋白分析的洗脱时间，同时保持筛查和确认所需的高灵敏度和分辨率。
该方法能够在12分钟内分离出45种以上常见的血红蛋白变体。这些包括巴氏胎儿血红蛋白、H、糖化血红蛋白A1C、罗利血红蛋白、霍普血红蛋白、I型血红蛋白、胎儿血红蛋白F、卡姆登血红蛋白、N - 巴尔的摩血红蛋白、I - 海威科姆血红蛋白、I - 巴黎血红蛋白、J - 巴尔的摩血红蛋白、N - 西雅图血红蛋白、格雷德血红蛋白、范宁 - 卢伯克血红蛋白、马尔默血红蛋白、南佛罗里达血红蛋白、A、芝加哥血红蛋白、G - 佐治亚血红蛋白、莱波雷 - 巴尔的摩血红蛋白、P - 加尔维斯顿血红蛋白、G - 库沙塔血红蛋白、莱波雷 - 波士顿血红蛋白、E、苏黎世血红蛋白、奥苏克里斯蒂安斯堡血红蛋白、A2、G - 费城血红蛋白、科尔勒布血红蛋白、拉斯血红蛋白、E - 萨斯卡通血红蛋白、里士满血红蛋白、D - 旁遮普血红蛋白（迪尔洛奇血红蛋白）、科隆血红蛋白、蒙哥马利血红蛋白、S、Q - 泰国血红蛋白、G - 圣何塞血红蛋白、A2'、哈沙龙血红蛋白、Q - 印度血红蛋白、坦帕血红蛋白、恒河泉血红蛋白、SG杂合血红蛋白、C - 哈莱姆血红蛋白、O - 阿拉伯血红蛋白、不列颠哥伦比亚血红蛋白和C型血红蛋白。该方法不仅能识别上述血红蛋白及其变体，还能准确测定它们的浓度，特别是Hb F和A2，分别对遗传性胎儿血红蛋白持续增多症和β地中海贫血的诊断很有用。
样品制备简单、方法分辨率高、血红蛋白浓度定量准确，再加上完全自动化，使其成为临床实验室常规诊断血红蛋白疾病的理想方法。
