Mutation Analysis Of The Spastin Gene In Chinese Patients With Hereditary Spastic Paraplegia Disease

Objective:To establish a technical platform in spastin gene diagnosis by combining polymerase chain reaction(PCR) with denaturing high-performance liquid chromatography(DHPLC) to detect the spastin gene mutation in Chinese patients with Hereditary Spastic Paraplegia(HSP or SPG) disease.To study the clinical manifestations of the patients in another 3 AD-SPG pedigrees and to find their relationship with spastin gene mutation.Methods:We detected the spastin gene mutation by DHPLC in probands of 10 SPG pedigrees and 18 sporadic cases.The abnormal amplifications detected by DHPLC were sequenced.Three AD-SPG family members were studied through clinical examinations,and then mutation analysis of the spastin gene was screened by PCR combined with DNA sequencing in the probands of these three AD-SPG pedigrees.Results:By using DHPLC in the mutation analysis of the spastin gene in probands of 10 SPG pedigrees and 18 sporadic cases,we found two abnormal peak forms in two patients.One was in exon 2 and another was in exon 11 of spastin,and the amplifications with abnormal peak form were sequenced.We found two point mutations in spastin:they were 651C→T(p.Gln144X) in exon 2 and 1617C→G (p.Leu466Val) in exon 11.All patients from the 3 AD-SPG pedigrees clinically manifested as classical SPG.No abnormal spastin gene mutation was detected in the probands of these 3 pedigrees. Conclusion:Applied the technology for mutation analysis of the spastin gene by DHPLC for the first time in China.Discovered two new point mutations in spastin, 651C→T(p.Gln144X) in exon 2 and 1617C→G(p.Leu466Val) in exon 11. Preliminarily tested the reliability for the mutation analysis by DHPLC in spastin.The 3 AD-SPG pedigrees had typical clinical manifesations of SPG.The pathogenesis had no association with mutation of the spastin gene.