title: Validation of a commercially available test that enables the quantification of the numbers of CGG trinucleotide repeat expansion in FMR1 gene.
abstract: In the present study, we evaluated a commercially available TP-PCR-based assay, the FastFraXTM FMR1 Sizing kit, as a test in quantifying the number of CGG repeats in the FMR1 gene. Based on testing with well characterized DNA samples from Coriell, the kit yielded size results within 3 repeats of those obtained by common consensus (n = 14), with the exception of one allele. Furthermore, based on data obtained using all Coriell samples with or without common consensus (n = 29), the Sizing kit was 97.5% in agreement with existing approaches. Additionally, the kit generated consistent size information in repeatability and reproducibility studies (CV 0.39% to 3.42%). Clinical performance was established with 198 archived clinical samples, yielding results of 100% sensitivity (95% CI, 91.03% to 100%) and 100% specificity (95% CI, 97.64% to 100%) in categorizing patient samples into the respective normal, intermediate, premutation and full mutation genotypes.
authors.affiliation: The BioFactory Pte Ltd, Singapore, Singapore.
authors.forename: Grace X Y
authors.identifier: http://orcid.org/0000-0002-4778-934X
authors.initials: GX
authors.lastname: Lim
authors.affiliation: The BioFactory Pte Ltd, Singapore, Singapore.
authors.forename: Minli
authors.initials: M
authors.lastname: Yeo
authors.affiliation: The BioFactory Pte Ltd, Singapore, Singapore.
authors.forename: Yvonne Y
authors.initials: YY
authors.lastname: Koh
authors.affiliation: Division of Human Genetics, Center for Biomedical Research, Faculty of Medicine, Diponegoro University, Semarang, Indonesia.
authors.forename: Tri Indah
authors.initials: TI
authors.lastname: Winarni
authors.affiliation: Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
authors.forename: Indhu-Shree
authors.initials: IS
authors.lastname: Rajan-Babu
authors.affiliation: Department of Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.|Khoo Teck Puat-National University Children's Medical Institute, National University Health System, Singapore, Singapore.|Department of Laboratory Medicine, National University Hospital, Singapore, Singapore.
authors.forename: Samuel S
authors.initials: SS
authors.lastname: Chong
authors.affiliation: Division of Human Genetics, Center for Biomedical Research, Faculty of Medicine, Diponegoro University, Semarang, Indonesia.
authors.forename: Sultana M H
authors.initials: SM
authors.lastname: Faradz
authors.affiliation: The BioFactory Pte Ltd, Singapore, Singapore.
authors.forename: Ming
authors.initials: M
authors.lastname: Guan
mesh_terms: D004247:DNA / Q000235:genetics*; D005260:Female; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics*; D005600:Fragile X Syndrome / Q000175:diagnosis* / Q000235:genetics; D005838:Genotype; D006801:Humans; D008297:Male; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D019680:Trinucleotide Repeat Expansion / Q000235:genetics*
chemical_list: D004247:DNA; D051860:Fragile X Messenger Ribonucleoprotein 1; C497683:FMR1 protein, human