Optimizing Tetra- Arms Pcr For The Detection Of Canton Mutation Inglucose-6-Phosphate Dehydrogenase Deficiency (G6pd Deficiency)

Abstract

Glucose-6-phosphate dehydrogenase deficiency is the most common hereditary enzymopathy in human and many G6PD mutations causing this disease have been found with different frequencies. Dealing with the concern caused by this abnormality is strongly dependent on precise detection and reliable screening of molecular tests. The aim of this study was the investigation of specificity and accuracy of Tetra primer ARMS PCR method comparing with conventional ARMS PCR. The main and primary task is to optimize the key components of Tetra- ARMS PCR for later improved performance. The size for genotyping of Canton mutation in G6PD deficiency is thirty samples including homozygote and heterozygote which were genotyped by ARMS PCR assay. After all the essential work done, results analysis and final DNA sequencing outcomes are two evidences to prove that Tetra-ARMS PCR is surpassing than conventional ARMS PCR. It means that both specificity and sensitivity of Tetra primer ARMS PCR are higher than ARMS-PCR. Comparing with Tetra primer ARMS PCR which represented 100% agreement with sequencing method, conventional ARMS PCR technique showed all disagreement, because of 4 discordant results. In conclusion, Tetra primer ARMS PCR method is a reliable and accurate technique and it is suggested that it can be used as a complementary method for diagnostic cases instead of conventional ARMS PCR method. This suggestion originated with perfect rate of agreement between outcomes of sequencing method, as a suitable method with rapid and cost benefits for detecting the mutations. For future research, this study will be useful for later accuracy improvement, sensitivity test and enhancement of Tetra-ARMS PCR.Hence the desire to develop Tetra-ARMS PCR to become the common molecular diagnostic method beside biochemical tests will be soon taken into practice. Keywords: G6PD Canton mutation Tetra-ARMS ARMS Optimization Validation