title: High-resolution melting analysis is a more effective approach for screening TSC genes mutations.
abstract: Tuberous sclerosis complex (TSC) is an autosomal dominant disorder with a prevalence of 1 in 95,136 in Taiwan. TSC is characterized by hamartomatous lesions in multiple organ systems. Genetic defects in TSC1 and TSC2 genes are the main causes of TSC. A molecular screening protocol using denaturing high-performance liquid chromatography (dHPLC) followed by DNA sequencing is currently performed to locate the genetic lesions in many clinical laboratories. The current screening approach is time consuming and inefficient. In this study, we analyzed all coding exons of TSC1 and TSC2 genes of 30 TSC patients and 47 unaffected family members using the traditional dHPLC protocol and our recently developed diagnostic platform based on high-resolution melting analysis (HRM) followed by bidirectional DNA sequencing. Data indicated that 20 mutations, including 5 mutations in TSC1 (2 sporadic, 1 familial mutation, and 2 of uncertain origin) and 15 mutations in TSC2 (14 sporadic and 1 familial mutation), 8 single-nucleotide polymorphisms (SNPs, including 3 SNPs found in irrelevant individuals without TSC phenotypes studied in the control group), and 3 variants with undetermined significance were identified, including 4 novel mutations. The sensitivities of HRM and dHPLC for TSC mutation screening were estimated as 95% and 75%, respectively. The specificities of HRM and dHPLC for TSC mutation screening were evaluated as 91% and 98%. In addition, results suggested our novel HRM screening protocol to be more economical. In conclusion, we successfully developed a superior approach for TSC genes mutation screening for clinical application.
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, Chang Gung University, 259 Wen-Hwa 1st Road, Kwei-Shan, Tao-Yuan, Taiwan, R.O.C.
authors.forename: Tz-Shiu
authors.initials: TS
authors.lastname: Tsai
authors.forename: Mu-Yi
authors.initials: MY
authors.lastname: Huang
authors.forename: Yu-Tsui
authors.initials: YT
authors.lastname: Chang
authors.forename: Chuan-Yu
authors.initials: CY
authors.lastname: Wang
authors.forename: Ju-Li
authors.initials: JL
authors.lastname: Lin
authors.forename: Po-Cheng
authors.initials: PC
authors.lastname: Hung
authors.forename: Shuan-Pei
authors.initials: SP
authors.lastname: Lin
authors.forename: Chien-Feng
authors.initials: CF
authors.lastname: Sun
authors.forename: Wei-Shun
authors.initials: WS
authors.lastname: Wang
authors.forename: Chung-Ming
authors.initials: CM
authors.lastname: Chang
authors.forename: Shih-Cheng
authors.initials: SC
authors.lastname: Chang
authors.forename: Da-Chang
authors.initials: DC
authors.lastname: Chu
mesh_terms: D002851:Chromatography, High Pressure Liquid; D005820:Genetic Testing; D006801:Humans; D009154:Mutation; D020641:Polymorphism, Single Nucleotide; D013624:Taiwan; D014402:Tuberous Sclerosis / Q000175:diagnosis* / Q000235:genetics*; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins / Q000235:genetics*
chemical_list: C000624650:TSC1 protein, human; C000624653:TSC2 protein, human; D000077004:Tuberous Sclerosis Complex 1 Protein; D000077005:Tuberous Sclerosis Complex 2 Protein; D025521:Tumor Suppressor Proteins