Ramos Cinthia, Ocampos Maristela, Barbato Ingrid Tremel, Graça Bicalho Maria da, Nisihara Renato
Post Graduate Program in Gynecology and Obstetrics, Universidade Federal do Paraná, Curitiba, Brazil.
Department of Genetics, Federal University of Paraná, Curitiba, Brazil.
Pract Lab Med. 2020 May 6;21:e00162. doi: 10.1016/j.plabm.2020.e00162. eCollection 2020 Aug.
Fragile X syndrome (FXS) is caused by expansion of the number of cytosine-guanine-guanine (CGG) repeats in the regulatory region of the gene (. The molecular diagnoses of FXS can be performed using two tests based on two different techniques, namely polymerase chain reaction (PCR) and Southern blotting (SB). However, both of these techniques have limitations. The purpose of this study was to evaluate the performance of the commercial FragilEase™ PCR kit for FXS diagnosis comparing to other laboratory methods.
and methods: This study had a retrospective design. We analyzed the performance of the FragilEase™ PCR kit using 90 DNA samples from patients with clinical suspicion of FXS or a family history of the syndrome using capillary electrophoresis and compared with the results obtained for the same samples using PCR, SB, and AmplideX PCR.
FragilEase™ PCR kit displayed high concordance with the results obtained using PCR, SB, and AmplideX PCR regarding the detection of normal, intermediate/gray zone, premutation, and full mutation alleles, as well as female homozygosity and mosaicism. The replicate sizes found using the FragilEase™ PCR assay varied on average by two CGG repeats.
FragilEase™ PCR, as well as other commercially available kits, efficiently detect mutations and simplify the workflow in laboratories that performing FXS diagnoses.
脆性X综合征（FXS）由基因调控区域中胞嘧啶-鸟嘌呤-鸟嘌呤（CGG）重复序列数量的扩增引起。FXS的分子诊断可通过基于两种不同技术的两项检测来进行，即聚合酶链反应（PCR）和Southern印迹法（SB）。然而，这两种技术都有局限性。本研究的目的是评估用于FXS诊断的商业FragilEase™ PCR试剂盒与其他实验室方法相比的性能。
本研究采用回顾性设计。我们使用毛细管电泳分析了来自临床怀疑患有FXS或有该综合征家族史患者的90份DNA样本，以评估FragilEase™ PCR试剂盒的性能，并将结果与使用PCR、SB和AmplideX PCR对相同样本获得的结果进行比较。
FragilEase™ PCR试剂盒在检测正常、中间/灰色区域、前突变和全突变等位基因以及女性纯合性和嵌合体方面，与使用PCR、SB和AmplideX PCR获得的结果高度一致。使用FragilEase™ PCR检测发现的重复序列大小平均相差两个CGG重复序列。
FragilEase™ PCR以及其他市售试剂盒能够有效检测突变，并简化进行FXS诊断的实验室的工作流程。
