Diagnosis of polyglutamine spinocerebellar ataxias by polymerase chain reaction amplification and Sanger sequencing

Chen,Changqiang; Fang,Xuqian; Sun,Shunchang

doi:10.3892/mmr.2018.9043

Diagnosis of polyglutamine spinocerebellar ataxias by polymerase chain reaction amplification and Sanger sequencing

Authors:
- Changqiang Chen
- Xuqian Fang
- Shunchang Sun
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Affiliations: Department of Laboratory Medicine, Ruijin Hospital North, Shanghai Jiao Tong University School of Medicine, Shanghai 201801, P.R. China
Published online on: May 22, 2018 https://doi.org/10.3892/mmr.2018.9043
Pages: 1037-1042

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Abstract

Spinocerebellar ataxia (SCA) is a group of genetic diseases of the nervous system with genetic and clinical heterogeneity. SCA is often caused by an expanded CAG repeat sequence in the encoding protein. Genetic testing is necessary to diagnose and classify the types of SCA. Next‑generation DNA sequencing usually generates a high error rate for insertion or deletion mutations, so it is unhelpful for classifying the types of SCA. In the present study, a Chinese SCA pedigree was preliminarily diagnosed with SCA1 using polymerase chain reaction (PCR) amplification. The propositus and his three younger siblings were diagnosed with SCA1 as a result of the identification of the length of the expanded CAG repeat sequence in the ATXN1 gene performed using Sanger sequencing. The current study presents a convenient and efficient method to identify causative mutations for polyglutamine SCA using PCR amplification followed by Sanger sequencing.

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1	Nibbeling EAR, Duarri A, Verschuuren-Bemelmans CC, Fokkens MR, Karjalainen JM, Smeets CJLM, de Boer-Bergsma JJ, van der Vries G, Dooijes D, Bampi GB, et al: Exome sequencing and network analysis identifies shared mechanisms underlying spinocerebellar ataxia. Brain. 140:2860–2878. 2017. View Article : Google Scholar : PubMed/NCBI
2	Dell'Orco JM, Pulst SM and Shakkottai VG: Potassium channel dysfunction underlies Purkinje neuron spiking abnormalities in spinocerebellar ataxia type 2. Hum Mol Genet. 26:3935–3945. 2017. View Article : Google Scholar : PubMed/NCBI
3	Sutton JR, Blount JR, Libohova K, Tsou WL, Joshi GS, Paulson HL, Costa MDC, Scaglione KM and Todi SV: Interaction of the polyglutamine protein ataxin-3 with Rad23 regulates toxicity in drosophila models of spinocerebellar ataxia type 3. Hum Mol Genet. 26:1419–1431. 2017. View Article : Google Scholar : PubMed/NCBI
4	Terada T, Kono S, Konishi T, Miyajima H and Ouchi Y: Altered GABAergic system in the living brain of a patient with spinocerebellar ataxia type 8. J Neurol. 260:3164–3166. 2013. View Article : Google Scholar : PubMed/NCBI
5	du Montcel Tezenas S, Durr A, Bauer P, Figueroa KP, Ichikawa Y, Brussino A, Forlani S, Rakowicz M, Schöls L, Mariotti C, et al: Modulation of the age at onset in spinocerebellar ataxia by CAG tracts in various genes. Brain. 137:2444–2455. 2014. View Article : Google Scholar : PubMed/NCBI
6	Li M, Pang SY, Song Y, Kung MH, Ho SL and Sham PC: Whole exome sequencing identifies a novel mutation in the transglutaminase 6 gene for spinocerebellar ataxia in a Chinese family. Clin Genet. 83:269–273. 2013. View Article : Google Scholar : PubMed/NCBI
7	Rothberg Gould BE and Rothberg JM: Massively parallel (‘next-generation’) DNA sequencing. Clin Chem. 61:997–998. 2015. View Article : Google Scholar : PubMed/NCBI
8	Miller SA, Dykes DD and Polesky HF: A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Res. 16:12151988. View Article : Google Scholar : PubMed/NCBI
9	Shakkottai VG and Fogel BL: Clinical neurogenetics: Autosomal dominant spinocerebellar ataxia. Neurol Clin. 31:987–1007. 2013. View Article : Google Scholar : PubMed/NCBI
10	Melo AR, Ramos A, Kazachkova N, Raposo M, Bettencourt BF, Rendeiro AR, Kay T, Vasconcelos J, Bruges-Armas J and Lima M: Triplet repeat primed PCR (TP-PCR) in molecular diagnostic testing for spinocerebellar ataxia type 3 (SCA3). Mol Diagn Ther. 20:617–622. 2016. View Article : Google Scholar : PubMed/NCBI
11	Scoles DR, Meera P, Schneider MD, Paul S, Dansithong W, Figueroa KP, Hung G, Rigo F, Bennett CF, Otis TS and Pulst SM: Antisense oligonucleotide therapy for spinocerebellar ataxia type 2. Nature. 544:362–366. 2017. View Article : Google Scholar : PubMed/NCBI
12	Lai S, O'Callaghan B, Zoghbi HY and Orr HT: 14-3-3 Binding to ataxin-1(ATXN1) regulates its dephosphorylation at Ser-776 and transport to the nucleus. J Biol Chem. 286:34606–34616. 2011. View Article : Google Scholar : PubMed/NCBI
13	McMurray CT: Mechanisms of trinucleotide repeat instability during human development. Nat Rev Genet. 11:786–799. 2010. View Article : Google Scholar : PubMed/NCBI
14	Chatterjee N, Lin Y, Santillan BA, Yotnda P and Wilson JH: Environmental stress induces trinucleotide repeat mutagenesis in human cells. Proc Natl Acad Sci USA. 112:3764–3769. 2015.PubMed/NCBI
15	Wang J, Shen L, Lei L, Xu Q, Zhou J, Liu Y, Guan W, Pan Q, Xia K, Tang B and Jiang H: Spinocerebellar ataxias in mainland China: an updated genetic analysis among a large cohort of familial and sporadic cases. Zhong Nan Da Xue Xue Bao Yi Xue Ban. 36:482–489. 2011.PubMed/NCBI