The Genetic Diagnosis Research On Glanzmann Thrombasthenia

Thrombasthenia ,also called Glanzmann thrombasthenia, which found by Glanzmann, is an autosomal recessive disorder. It commonly found in consanguineous mating people and no significant difference in male or in female. This disorder caused by the mutant of ITGA2B and ITGA3 which leading the quantitative or qualitative abnormality of platelet glycoprotein IIb—IIIa receptor(GP IIb/ IIIa).Objective: To study the pathogenic locus and mechanism of Glanzmann thrombasthenia , we diagnose a GT pedigree with a efficient, rapid method.Methods: Extracted peripheral blood RNA from the patients with GT , we used RT-PCR to amplify the full length of ITGA2B and ITGA3. After sequencing, the restriction enzymes and picked a single colony were used to analysis the mutants of the genes. Then, extracted peripheral blood DNA , amplification and DNA sequencing evidence the locus of this mutants.Result: To amplify and sequencing-analysis the cDNA of GT patients, we found a deletion locus on ITGA2B 508-607 which leading the absence of CDS160 - CDS192. The Complex mutants of +72 in EXON4(C→G) and +5 in INTRON4(G→C) was found through genomic DNA analysis. The new exceptional case ,which had abnormal RNA splicing caused by mutant ,was not found in Human Gene Mutation Database(HGMD) and Glanzmann thrombasthenia database.Conclusion: To analysis Glanzmann thrombasthenia, the ITGA2B and ITGA3 mRNA should be diagnose before routine gene diagnosis. In comparison with direct-sequencing individual exons, this method was not only benefit at cost and speed, but also avoidance the effect of abnormal transcription caused by DNA mutants.