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: A Single Common Assay for Robust and Rapid Fragile X Mental Retardation Syndrome Screening From Dried Blood Spots

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: A Single Common Assay for Robust and Rapid Fragile X Mental Retardation Syndrome

Screening From Dried Blood Spots

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: a. Nama Jurnal Frontiers in Genetics

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A single common assay for robust and rapid fragile X mental retardation syndrome screening from dried blood spots (Article) (Open Access)

, , , ,

Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, SingaporeKhoo Teck Puat – National University Children's Medical Institute, National University Health System, Singapore,

SingaporeDivision of Human Genetics, Center for Biomedical Research, Faculty of Medicine, Diponegoro University, Semarang,

Indonesia

AbstractBackground: FMR1 CGG trinucleotide repeat hyper-expansions are observed in 99% of individuals with fragile Xmental retardation syndrome (FXS). We evaluated the reliability of a rapid single-step gender-neutral molecularscreen for FXS when performed on DNA isolated from dried blood spots. Methods: DNA was extracted from driedblood spots of 151 individuals with intellectual disability or autism spectrum disorder , whose FMR1 repeatgenotypes are known. Dried blood spots were blinded prior to DNA extraction and analysis by triplet primed PCR (TP-PCR) and melt curve analysis (MCA). All expansion-positive and representative expansion-negative samples were alsogenotyped by fluorescent TP-PCR and capillary electrophoresis (CE) to confirm repeat expansion status. Results: Threemales and 12 females were classified as expanded by TP-PCR MCA, and were subsequently sized by fluorescent TP-PCR CE. Two males and four females carried premutations, while one male and eight females carried full mutations.All 19 non-expanded samples that were sized were confirmed as carrying only normal alleles. Replicate analysis ofrepresentative expansion-positive samples yielded reproducible melt peak profiles. TP-PCR MCA classifications werecompletely concordant with FMR1 CGG repeat genotypes. Conclusion: TP-PCR MCA of dried blood spot DNAaccurately and reliably identifies presence/absence of FMR1 CGG repeat expansions in both genders simultaneously.This strategy may be suitable for rapid high-throughput first-tier screening for fragile X syndrome . Copyright© 2018 Tan, Lian, Faradz, Winarni and Chong. This is an open-access article distributed under the terms of theCreative Commons Attribution License (CC BY).

Author keywordsDried blood spot FMR1 Fragile X syndrome Melt curve analysis (MCA) Trinucleotide repeat

Triplet-primed PCR (TP-PCR)

Funding details

Funding sponsor Funding number Acronym

University of the Punjab 176-13/UN7.5.1/PG/2016 PU

Temple University Health System NUHSRO/2015/056/STB/BBP FY15/01 TUHS

Funding textThis work was funded in part by a PUPT Kemenristek Dikti Grant (No. 176-13/UN7.5.1/PG/2016) to TW and a NationalUniversity Health System grant (NUHSRO/2015/056/STB/BBP FY15/01) to SC.

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Open AccessFrontiers in GeneticsVolume 9, 2018, Article number 582

Tan, V.J.a Lian, M.b Faradz, S.M.H.c Winarni, T.I. Chong, S.S.a,b,d

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ISSN: 16648021Source Type: Journal

DOI: 10.3389/fgene.2018.00582Document Type: Article

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paecs@nus.edu.sg

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