全自动高效液相色谱分析系统在地中海贫血诊断中的应用

摘要

Objective To evaluate the application of high-performance liquid chromatography (HPLC) in diagnosis and screening of thalassemia. Methods Automated HPLC was used to measure HbF and HbA2 in 100 genetically diagnosed thalas-semic patients and 35 normal children. The results were compared with those from traditional tests including alkali denaturation test and cellulose acetate electrophoresis. The diagnose accordance rates, sensitivity and specificity were compared. Results Seventy-fourβthalassemia, 64 were heterozygous with single mutations and 10 were compound heterozygous with double muta-tions. Twenty-sixαthalassemia, 25 were compound mutations and one was heterozygous with single mutation. The HbF percent-age from HPLC was higher than that from alkali denaturation tests in either thalassemia or normal children (P<0.01). HbF level from HPLC inα-thalassemia was signiifcantly different from that in the normal children (P=0.011). The percentage of HbA2 from HPLC was higher than that from cellulose acetate electrophoresis (P=0.010). HbA2 in the single heterozygousβ-thalassemia were twice higher than that in the double heterozygous mutatedβ-thalassemia (P<0.01). The combination of HbF-HbA2 (≥4.0%) from HPLC with MCV (<80 lf) and MCH (<27 pg) had high accordance rates (99.3%), sensitivity (99.0%) and speciifcity (100.0%) in diagnosis of thalassemia. Conclusions When the results of HPLC are combined with MCV and MCH, it can be applied to the diagnosis of thalassemia with high speciifcity, high sensitivity and has high diagnostic accordance rate with genetic results. HPLC can be an ideal approach to screenβthalassemia.%目的：探讨全自动高效液相色谱（HPLC）分析检测系统在地中海贫血(地贫)诊断和筛查中的临床应用价值。方法入选100例基因诊断确诊地贫患儿及35例正常儿童，应用HPLC检测系统以及传统碱变性法、醋纤电泳法定量分析HbF和HbA2；并比较不同检测方法与相应β地贫基因诊断结果的符合率、敏感性与特异性。结果β地贫74例，64例单杂合子，10例复合杂合子；α地贫26例，1例单杂合子，25例复合杂合子。在地贫患儿和正常儿童中，HPLC法测定的HbF百分含量均高于碱变性法，差异有统计学意义（P<0.01）。HPLC法测得的HbF在α地贫患儿和正常儿童间的差异亦有统计学意义（P=0.011），对α地贫的分辨力高于碱变性法。α地贫患儿，HPLC法测得的HbA2百分含量高于醋纤电泳法，差异有统计学意义（P=0.010）；β地贫患儿，HPLC法和醋纤电泳法测得的单杂合子的HbA2含量均高于复合杂合子，差异有统计学意义（P<0.01）。HPLC方法测定HbF-HbA2（≥4.0%）联合MCV（<80 lf）、MCH（<27 pg），与α或β基因诊断的符合率达99.3%，灵敏度和特异度分别为99.0%和100.0%。结论采用HPLC法检测HbA2和HbF，并结合MCV、MCH，与基因诊断结果的符合率、灵敏度和特异度均高，可成为大规模筛查β地贫的一种理想方法。