Nested Polymerase Chain Reaction For Detection Of Spasmodic Torticollis Dopamine D5Receptor Gene Mutation

Background:Dystonia occurs with dopamine,5-serotonin and acetylcholineneurotransmitter disorders. Onset spasmodic torticollis may or dopaminetransporter gene polymorphism D5receptors. D5receptor gene microsatellitepolymorphism studies of patients with spasmodic torticollis found4allelefrequency is higher than the control group, suggesting that D5receptormutation may be predisposing factors spasmodic torticollis. The D5receptordysfunction by affecting the basal ganglia dopaminergic pathway, affectingbasal ganglia function, leading to an increase in activity, indicating thatspasmodic torticollis the pathogenesis of the dopamine system abnormalitiesmay have a certain relationship.Preliminary study to explore the correlationbetween dopamine D5receptor gene polymorphism and spasmodic torticollis，Found877,1096,1199,1229,1268,1269sites appear on the D5dopaminereceptor gene (DRD5gene) mutation ORF single nucleotide differencesbetween sites, which were statistically significant, may be inclined with spasticneck susceptibility. Objective: Verify that the mutant gene on DRD5spasmodic torticollis found inprevious studies of ORF877,1096,1199,1229,1268,1269sites whether theincidence of spasmodic torticollis correlation further as spasmodic torticollismolecular biology provides the basis for treatment.Methods: Single nucleotide differences appear locus mutation according to theprevious study in patients with spasmodic torticollis found ORF877,1096,1199,1229,1268,1269sites at six sites within the external design, theouter primers nested PCR amplification，Nested PCR amplification of the targetgene.After nested PCR amplified target gene gene sequencing, ultimatelydetermine the circumstances of each sample point mutation.Results:After gene sequencing detected the presence of single nucleotidemutations of genes on ORF DRD5877points, the A variant of G. Consistentwith previous findings.The ORF1096,1199,1229,1268,1269sites withoutmutations. ORF on a877-point single-nucleotide mutations obtained bystatistical analysis of p <0.05considered statistically significant. The site OR=4.833>1, more common in spasmodic torticollis group, suggesting that thislocus may be a risk factor for spasmodic torticollis.Conclusion：1.1096,1199,1229,1268,1269on ORF DRD5gene locus without mutation.2.877single nucleotide differences between the mutant locus of points on ORFDRD5statistically significant genes, this locus may be associated withspasmodic torticollis about possible risk factors for spasmodic torticollis.