应用HPLC检测血红蛋白Constant Spring复合血红蛋白H病

Apllication of HPLC for diagnosis of HbCS/HbH diseases

目的探讨应用高效液相色谱技术(HPLC)快速诊断血红蛋白(constant spring,HbCS)复合HbH病的可行性,并分析其血液学特点。方法采用HPLC方法分析312例经PCR方法诊断为东南亚缺失型α地中海贫血患儿的各血红蛋白组分,对HbCS复合HbH病进行非基因检测,同时采用RDB方法检测312例患儿的HbCS基因突变,并测定其相关的血液学指标。结果①应用HPLC方法检出23例(7.37%)HbCS复合HbH病,HbCS复合HbH病患儿既可见一HbH峰,又可见一HbCS峰;采用RDB方法检出24例(7.69%)患儿有HbCS基因突变,HPLC方法与分子生物学方法对HbCS复合HbH病诊断的一致率达99.7%。②与其他288例东南亚缺失型α地中海贫血患儿相比,HbCS复合HbH病患儿的红细胞计数(RBC)、血红蛋白(Hb)、红细胞平均体积(MCV)、平均红细胞血红蛋白量(MCH)和红细胞平均血红蛋白浓度(MCHC)值均偏低(P<0.01)。结论①HPLC方法诊断HbCS复合红蛋白H病具有比较高的准确度,在没有开展基因诊断的情况下,可作为快速诊断HbCS复合HbH病的方法进行推广。②HbCS复合HbH病患儿比东南亚缺失型α地中海贫血患儿有更严重的血液学表现。

[Objective] To evaluate the usefulness of high performance liquid chromatography （HPLC） as a diagnosis method for HbCS/HbH disease and analyzed the characterization of hematological parameters. [Methods] In the 312 cases of SEA-α-thalassemia children diagnosed by polymerase chain reaction （PCR）, the precentages of homoglobin fractions were detected by HPLC. At the same time, reverse dot blot （RDB） were performed to determine HbCS gene mutations respectively and determined the hematological parameters of 312 cases. [Results] In the 312 cases of SEA-α-thalassemia children, 23 cases of HbCS/HbH disease （7.37%） were diagnosed by HPLC. In the HbCS/HbH disease child, a HbH peak and a HbCS peak were pre- sent in the ehromatogram. At the same time, 24 cases of HbCS/HbH disease （7.69%） were diagnosed by RDB, the agreement rate of HPLC and RDB was 99.7%. HbCS/HbH Disease children have significantly lower RBC, Hb, MCV, MCH and MCHC than those of 288 cases of SEA-ct-thalassemia （both P〈0.01）. [ Conclu- sion] HPLC is suitable for diagnosis of HbCS/HbH Disease, HPLC can be popularized as a rapid diagnosis method of HbCS/HbH disease. The children with HbCS/HbH disease presented with more severe hematological phenotypes.