Development of loop-mediated isothermal amplification (LAMP) combined with a lateral flow dipstick DNA biosensor for detection of α thalassemia 1 [Southeast Asia (SEA), --SEA deletion]

Abstract

This study aimed to develop a one-step loop-mediated isothermal amplification combined with a lateral flow dipstick (LAMP-LFD) DNA biosensor for the detection of α 0 -thalassemia (SEA deletion). Genomic DNA was obtained from 48 α 0 -thalassemia (SEA deletion)-positive and 48 (SEA deletion)-negative (normal) Thai subjects. DNA was analyzed by LAMP assay at 61°C for 60 min using specific primers for the α 0 -thalassemia (SEA deletion) type. Following amplification, the products were visually detected using an LFD, with results available within 5–10 min. The one-step LAMP-LFD DNA biosensor could detect DNA carrying the α 0 -thalassemia (SEA deletion) type at levels as low as 200 pg, with 100% sensitivity and 100% specificity. Furthermore, no cross-reactivity was observed with DNA carrying other α 0 -thalassemia deletion types (THAI deletion). The LAMP-LFD DNA biosensor is useful for identifying carriers of α 0 -thalassemia (SEA deletion) (the most common deletion type found in Thailand and across Southeast Asia). This novel one-step LAMP-LFD kit will be beneficial for medical laboratory technologists performing genetic carrier screening tests as it is simple to perform and requires minimal equipment and time. Furthermore, this method supports clinicians in providing genetic counseling and implementing preventive strategies against severe thalassemia, a significant public health concern in Thailand.