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一步法制备荧光开启传感器检测BRCA2

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筛查乳腺癌易感基因2(BRCA2)基因突变对于乳腺癌患病风险评估、发病检测、早期诊断及基因治疗均具有重要临床意义.现有乳腺癌基因传感检测方法大多需要复杂的处理过程,成本较高且对单碱基突变识别能力较弱.为简化基因传感器制备过程,提高其应用可能性,文章通过DNA分子发夹结构设计,一步实现了 BRCA2的检测.其中分子发夹茎部含BRCA2互补序列,茎与环连接处标记四甲基罗丹明荧光基团(TAMRA),环部5个G碱基通过光诱导电子转移猝灭部分荧光信号,降低背景荧光信号;待测BRCA2序列与发夹茎部序列竞争结合,发夹结构被打开,荧光信号上升,测量上升的荧光信号强度即可实现一步法定量检测靶基因.优化实验结果表明,分子发夹浓度不同时,传感器检测范围不同.当分子发夹浓度为150 nmol·L-1时,对BRCA2的线性检测区间为1~50 nmol·L-1;浓度为300和600 nmol·L-1时,线性检测区间分别为1~100和1~200 nmol·L-1.三种浓度传感器检出限分别为0.54、0.72和1.81 nmol·L-1.此外,传感器对BRCA2的特异性检测较为突出,尤其对单碱基突变的识别能力较强.该方法制备与检测过程简单且特异性突出,不仅可用于乳腺癌高危人群的早期筛查,还可拓展应用于其他种类的基因检测,为基因传感器的推广应用提供一种制备简单、操作便捷的新方法.
One-Step Detection of BRCA2 by Fluorescence Turn-On Sensor
Screening for BRCA2 gene mutations is clinically important for breast cancer risk assessment,morbidity detection,early diagnosis and gene therapy.Most existing breast cancer gene sensing assays require complex processing,high cost and weak recognition of single-base mutations.To simplify the process of gene-sensor preparation and improve its application possibility,the article realized the detection of BRCA2 in one step through the design of DNA molecular hairpin structure.The molecular hairpin stem contains BRCA2 complementary sequences,and the stem-loop junction is labeled with a tetramethylrhodamine fluorescent group.The target gene can be measured in one step.The optimized experimental results showed that the sensor's detection range was different when the concentration of molecular hairpin was different.The linear detection range of BRCA2 was 1~50 nmol·L-1 at 150 nmol·L-1,1~100 and 1~200 nmol·L-1 at 300 and 600 nmol·L-1,respectively,and the detection limits for the three concentrations were 0.54,0.72,and 1.81 nmol·L-1,respectively.In addition,the specificity of the sensor for detecting BRCA2 was outstanding,especially for identifying single-base mutations.The method is simple and specific and can be used not only for early screening of breast cancer in high-risk groups but also for other types of genetic tests,providing a new method with simple preparation and convenient operation for applying genetic sensors.

Breast cancer susceptibility gene 2One-step methodFluorescence turn-on sensorGene detection

任林娇、魏铭航、陈青华、张培、闫艳霞、薛梦晓、秦自瑞、姜利英

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郑州轻工业大学电气信息工程学院,河南郑州 450002

郑州轻工业大学计算机与通信工程学院,河南郑州 450002

郑州轻工业大学量子科技研究院,河南郑州 450002

乳腺癌易感基因2 一步法 荧光开启传感器 基因检测

国家自然科学基金国家自然科学基金中原科技创新领军人才项目河南省科技攻关计划河南省科技攻关计划河南省科技攻关计划河南省科技攻关计划

6207329912004348224200510026212102210196212102210002212102210278232102220008

2024

光谱学与光谱分析
中国光学学会

光谱学与光谱分析

CSTPCD北大核心
影响因子:0.897
ISSN:1000-0593
年,卷(期):2024.44(6)
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