Objective: To explore the application value of high performance liquid chromatography (HPLC) and erythrocyte parameters for screening of thalassemia in Shanghai. Methods: The hemoglobin of 1 029 samples from patients with suspected thalassemia and with suspected hemolytic anemia were analyzed using HPLC. Genetic analysis of the globin were performed simultaneously with GAP-PCR,reverse dot blot(RDB) and DNA sequencing. Results: Among 1029 samples, 591（57.43%） were diagnosed as having thalassemia and 14 (1.36%) having structural hemoglobin variants.Compared with results of genetic analysis,the sensitivity,specificity,positive predictive value and negative predictive value of HbA2 >3.9% by HPLC as cut-off value for diagnosing β-thalassemia were 95.84%, 96.87%, 95.40%, 97.20%,with AUC of 0.962. When HbA2≤2.1% by HPLC as the cutoff value combined with presence of rapid band were taken for diagnosing α-thalassemia moderate（HbH disease）, sensitivity, specificity, positive predictive value, negative predictive value were 96.32%, 52.60%, 100.00 %, with AUC of 0.990. If f HbA2 ≤3.2% by HPLC was taken as the cutoff value for diagnosing minor α-thalassemia or α-thalassemia trait, the corresponding indice were 97.76%, 53.11%, 24.0%, 99.4%,and AUC was 0.753. If MCV≤73.3 fl,MCH≤23.5 pg,MCHC≤324g/L were taken as the cutoff value alone for diagnosing thalassemia, sensitivity and specificity were 87.98%,77.51%, 93.73% and 69.86%, 89.20%,47.61%,respectively. Conclusions: Based on RBC parameters analysis,results of HPLC technology have a good consistence with the resluts of gene detection in the diagnosis of β-thalassemia, HbH disease and structural hemoglobin variant, while a low specificity and positive predictive value may occur in the diagnosis of minor α-thalassemia orα-thalassemia trait.