Development Of Non-deletion Alpha Thalassemia Genotyping Detecting Kit

Alpha-thalassemia is one of the most common human monogenic hereditary diseases inthe World, including southern China. Generally,-thalassemia is classified as deletional ornon-deletional according to the mutational mechanism involved. The deletion types comprisethe majority of cases of-thalassemia, but the non-deletion types also got a ample proportions.There is no suitable product to diagnose Non-deletion alpha thalassemia(T) at present. It isnecessary to develop a kit for qualitative analysis of Non-deletion alpha thalassemia mutationin Chinese. On the base of PCR-Reverse Dot Blot, specific PCR primers and probes for thethree nondeletion-thalassemia were designed. The PCR reaction systern, StripAssay hybridsystem and color systern were set up and the conditions were optimized.Biotin labeled primers Bio-ATF2and Bio-ATR1amplified exon III of2genespecifically. The hybridization between this300bp fragment and the probes can identify threetypes of the Chinese common Non-deletion thalassemia under the appropriate conditions. In28cases, StripAssay typing was completely concordant with DNA sequence. The detectionsof other genotypes were all negative. Besides, the kit can reach the minimum detection limitof2ng/μL. It could get the same repeatable and stable result under certain condition.The kit comprises PCR reaction system, StripAssay hybrid system and color system.3types of Non-deletion-thalassemia commonly encountered in Chinese can be identifiedqualitatively, and got the identical genotype as DNA sequence. The StripAssay proved to be afast, easy-to-perform, high sensitive and specific method to diagnose Non-deletionalthalassemia. It will reduce the missing rate of thalassemia diagnosis, and cut down theburden on society and family caused by Non-deletion alpha thalassemia patients.