HPLC and gene analysis to diagnose thalassemia

Objective To elevate the value of HPLC and gene analysis for diagnosis of thalassemia.Methods 174 blood samples with MCV80fl and MCH26pg were collected from 83 males and 91 females,the age ranging from 5 months to 77 years.The content of HbA2 was detected by HPLC.After DNA extraction,amplification by PCR,the common types of alpha-thalassemia(SEA,-α3.7,α4.2) were assayed by electrophoresis.The common types of beta-thalassemia(CD41-42M,-28M,IVS-II-654M,CD71-72M) were detected by reverse dot blot(RDB).Results Among the 174 samples,27 had a normal content of HbA2 and 147 abnormal.106 cases(60.9%) of HbA2≤2.5%,were suspected of alpha-thalassemia and 41 cases(23.6%) of HbA2≥3.5% were suspected of beta-thalassemia.Gene analysis were used to test the abnormal cases,in whom,66 cases(37.9%) were confirmed as alpha thalassemia,34 cases were normal,and 5 cases were not be clarified.Conclusion There exists discrepancy in diagnosing alpha-thalassemia by HPLC or gene analysis,but for beta-thalassemia,they are coincidence.HbA2 is not related to thalassemia.HPLC is suitable for diagnosing beta-thalassemia.For alpha-thalassemia,gene analysis should be used to raise the accurate rate of diagnosis.