The ATP1A2 Gene Mutation Detetion Of A Alternating Hemiplegia Of Childhood Family

Background Alternating hemiplegia of childhood (AHC) is a rare syndrome, characterised by repeated onset of episodic hemi-or quadriplegia, accompanied with tonic-dystonic abnormal posture,paroxysmal nystagmus and ophthalmopl-egia;progressive dysgnosia is an important feature of AHC. Theaetiological agent of AHC is still not very clear. rencent research in molecular have got certain progress. A researsh have found that a ATP1A2 gene coding for the a2 subunit of the Na, K pump, associated with familial hemiplegic migraine, may lead to AHC; We found a novel heterozygous mutation segregating (1237C→A) in the hydrolase domain with the disease and causing a threonine to asparagine replacement (T378N).This is the first mutation associated with AHC identified so far. Currently there is noeffective treatment methods about AHC, we try to find the gene mutations described in the literature or new genetic mutations,which is important to discover new clinical treatments.Objective Crrry the ATP1A2 gene mutation detection of a alternating hemiplegia of childhood family, in order to find new genetic mutation leading to AHC. Methods Extract 5ml blood from 5 patient of a AHC family, then use EDTA for anti-freezing, store in-20℃。Extract genomic DNA from the blood sample according to the operation steps, detect which absorbance od260/od280>1.8.Design the primer according to the mutation domain by Primer5(primer sequence forward 5-GGTCTAGGGTAAGGTTATGG-3'; reverse 5'-CCTCCTCTAATCCTATCCAC-3'),PCR (reaction system 20ul; circulation conditions:degeneration 95℃5min, degeneration 94℃30s, renaturation 57℃30s, extension 72℃30s, termination 72℃7 min,30cricle); Carry 2% agarose gel electrophoresis to detect the molecular weight of PCR product with DL2000 as relative molecular marks and (?)-actin as beta-actin.Repeat PCR with PCR product and primer to get plenty of DNA samples, each packing,send to the boshang company for gene sequencing.Conclusion After gene sequencing, we compares normal ATP1A2 gene sequences with the gene sequencing sequence,yet no mutation have been found.there are no obvious doublet in the gene sequencing figure.there are no gene mutation being found in all 5 patient DNA samples.