Combined use of gap-PCR and next-generation sequencing improves thalassaemia carrier screening among premarital adults in China

Aims Thalassaemia is one of the most common genetics disorders in the world, especially in southern China. The aim of the present study was to investigate the feasibility of combining the gap-PCR and next-generation sequencing (NGS) for thalassaemia carrier screening in the Chinese population. Methods Blood samples were obtained from 944 prepregnancy couples; thalassaemia carrier screening was performed by using a routine haematological method and a combination of gap-PCR and NGS method. Results We found that the alpha thalassaemia carrier rate was 11% (207/1888); the beta thalassaemia carrier rate was 3.7% (70/1888); the composite alpha thalassaemia and beta thalassaemia carrier rate was 0.4% (8/1888). We also identified seven novel mutations, including HBA1: c.412A>G, -50 (G>A), HBB: c.*+129T>A, HBB: c.-64G>C, HBB: c.-180G>C, HBB: c.*+5G>Aand HBB: c.-113A>G. By comparing the combined gap-PCR and NGS method, the MCV+MCHand HbA2 detection strategy showed a lower sensitivity of 61.05% (105/172) and a higher missed diagnosis ratio of 38.95% (67/172) for alpha thalassaemia mutations. The sensitivity was improved with the MCV+MCHand HbA2 detection screen when compared with MCV+MCH detection for beta thalassaemia (98.51% vs 85.90%). Conclusions Our study suggests the combined gap-PCR and NGS method is a cost-effective method for the thalassaemia carrier screening, particularly for the alpha thalassaemia mutation carriers.