DNA biosensor paper for molecular detection of β-thalassemia

Thalassemia is one of the most common single gene disorders and considered to be a major public health problem. It occurs by a point mutation in the HBB gene. The molecular detection of β-thalassemia is necessary for prenatal molecular diagnosis. At present, Most of clinical diagnostic methods are based on polymerase chain reaction.Therefore, due to the benefits of prenatal genetic screening, the use of sensitive, low cost, rapid methods for the clinical diagnostic is valuable [1, 2]. The purpose of this study is developed method for Simultaneous detection of six frequent β-globin mutations in Khuzestan population.The method is based on color changes and the naked eye detection upon enzymatic (AP) activity after specific amplification of the globin gene and hybridization target DNA with the allele specific oligonucleotide probes on nylon paper. The effect of important parameters, such as concentration of probe, incubation time and temperature, type and volume of buffer were investigated and optimized.This method due to design specific probe for each mutation, has high selectivity and no signal background, false positive and false negative signal. Under the optimum conditions, detection limit was 70 ng genome per sample. The method was successfully applied to diagnosis of β-thalassemia mutation in blood, amniotic fluid and Chorionic villus samples. This StripAssay proposes fast and reliable diagnostic method for prenatal screening.