A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene

作者： Huilin Wang*1, Xiaofan Zhu*2,3, Baoheng Gui*3,4, Wan Chee Cheung2, Mengmeng Shi2,3, Zhenjun Yang2,3, Ka Yin Kwok2, Ricky Lim5, Sanna Pietilä5, Yuanfang Zhu6,7, Kwong Wai Choy2,3 1Central Laboratory, Bao'an Maternity and Child Health Hospital,Jinan University, 2Department of Obstetrics and Gynaecology,The Chinese University of Hong Kong, 3Shenzhen Research Institute,The Chinese University of Hong Kong, 4Department of Genetics and Metabolism,Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, 5PerkinElmer Diagnostics, 6Department of Obstetrics and Gynecology, Bao'an Maternity and Child Health Hospital,Jinan University, 7Maternal-Fetal Medicine Institute, Bao'an Maternity and Child Health Hospital,Jinan University

简介：

Overview

This article presents a cost-effective polymerase chain reaction-based assay for quantifying cytosine-guanine-guanine trinucleotide repeats in the Fragile X mental retardation-1 gene. The method enhances molecular diagnosis and screening of Fragile X syndrome and related disorders with improved turnaround time and reduced equipment investment.

Key Study Components

Area of Science

• Molecular Biology
• Genetics
• Clinical Diagnostics

Background

• Fragile X syndrome is a genetic condition caused by mutations in the FMR1 gene.
• Traditional diagnostic methods can be time-consuming and costly.
• There is a need for efficient and reliable screening techniques.
• This study introduces a PCR-based assay to address these challenges.

Purpose of Study

• To develop a robust assay for quantifying trinucleotide repeats in the FMR1 gene.
• To facilitate the classification of Fragile X syndrome and related disorders.
• To reduce the time and cost associated with molecular diagnosis.

Methods Used

• Preparation of PCR buffer mix and DNA samples.
• Thawing of reagents at room temperature.
• Vortexing and spinning down samples before use.
• Application of the PCR assay to classify FXS and related disorders.

Main Results

• The assay demonstrated robustness in detecting various mutations.
• Results were reported rapidly, enhancing clinical decision-making.
• It effectively classified the full spectrum of Fragile X-associated disorders.
• The method proved to be cost-effective with minimal equipment requirements.

Conclusions

• The PCR-based assay is a significant advancement in Fragile X diagnostics.
• It offers a reliable and efficient alternative to traditional methods.
• This approach can improve patient outcomes through timely diagnosis.

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