论文标题
高灵敏度电化学DNA传感器，用于检测FFPE样品中体细胞突变
High sensitivity electrochemical DNA sensors for detection of somatic mutations in FFPE samples
论文作者
论文摘要
我们提供了新的高性能标签的非瓦拉达型电化学DNA传感器。 Langmuir-Blodgett（LB）沉积技术基于结晶碳纳米管（CNT）阵列的平台的DNA传感器。 CNT阵列悬挂在纳米多孔阳极氧化铝（氧化铝，AOA）上。单链（SS）19-和20-碱基寡核苷酸和双链（DS）核苷酸序列被用作所选分子靶标的探针 - 人Kras（Kirsten大鼠肉瘤病毒癌基因同源性）基因。从健康供体和FFPE（福尔马林固定，石蜡床）样品的胎盘中分离出基因组脱氧核糖核酸（DNA）。拉曼光谱分析和电化学介电光谱法用于对靶DNA进行测序。光学和电化学检测（DNA测序的变体）是基于在DNA同型形成后生长的筛选效果。我们证明了此类技术非常敏感，并允许检测attomolar DNA浓度，较少。结果，KRAS外显子2，密码子12，c.35g $> $ $ $ $ a突变在与FFPE结直肠癌肿瘤组织样品中分离的人类基因组DNA中成功歧视。
We offer new high-performance label-free electrochemical impedimetric DNA sensors of non-faradaic type. The DNA sensors based on a platform of crystalline carbon nanotube (CNT) arrays are fabricated by the Langmuir--Blodgett (LB) deposition technique. The CNT arrays are suspended on a nanoporous anodic alumina (aluminium oxide, AOA) support. Single-stranded (ss) 19- and 20-base oligonucleotides and double-stranded (ds) nucleotide sequences were used as probes for chosen molecular target -- human KRAS (Kirsten Rat Sarcoma viral oncogene homolog) gene. Genomic deoxyribonucleic acids (DNAs) were isolated from placenta of healthy donors and FFPE (formalin-fixed, paraffinembedded) samples of tumor tissue. Raman spectral analysis and electrochemical dielectric spectroscopy were used to sequence the target DNAs. The optical and electrochemical detection (variants of DNA sequencing) were based on a screening effect that grows after the DNA homoduplex formation. We demonstrated that such technologies are very sensitive and allow to detect attomolar DNA concentrations and less. As result, the KRAS exon 2, codon 12, c.35G$>$A mutation was successfully discriminated in human genomic DNA isolated from FFPE colorectal cancer tumor tissue samples.
