Accurate and rapid detecting β thalassemia by denaturing high-performance liquid chromatography / 中华检验医学杂志

ABSTRACT

Objective To evaluate the application value of denaturing high-performance liquid chromatography (DHPLC) as a rapid gene typing tool for β thalassemia. Methods 226 suspicious samples were screened with MCV, RDW, erythrocytcte agility and hemoglobin electrophoresis. The final diagnosis ofβ thalassemia genotype was made by DHPLC and PCR-reverse dot blot (PCR-RDB). Results Sixty-nine samples (30. 5% ) were eventually diagnosed as βthalassemia by PCR-RDB. The genotyping results for βthalassemia identified by DHPLC were complete agreement with genotyping results by PCR-RDB. We found 37 cases of CD41/CD42 ( - TCTT) frame shift mutation(54% ) ; 12 cases of IVS - Ⅱ - 654 (C→T) insertion mutation( 17% ) ;10 cases of TATA - 28 (A→G) transcription mutation ( 15% ) ;5 cases of CD17 (A→T)nonsense mutation ( 7% ) ; 5 cases of CD71/CD72 ( + A) frame shift mutation (7%). Conclusion The DHPLC is a rapid, sensitive , efficient and highly accurate assay in the diagnosis of β-thalassemia.