Naked-eye detection of nucleic acids through rolling circle amplification and magnetic particle mediated aggregation
Naked-eye detection of nucleic acids through rolling circle amplification and magnetic particle mediated aggregation
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DOI：
10.1016/j.bios.2013.03.056
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发表时间：
2013-09-15
影响因子：
12.6
通讯作者：
Fang, Qun
中科院分区：
文献类型：
--
作者：
Lin, Caiqin;Zhang, Yunxia;Fang, Qun
我们提出了一种通过等温扩增和磁性粒子介导的聚集进行序列特异性和裸眼检测核酸（NA）的新方法。挂锁探针设计用于与目标基因上的特定序列杂交，然后进行连接和滚环扩增（RCA）。然后将磁性颗粒 (MP) 添加到 RCA 产品溶液中。 MP 会与长线性 DNA 线圈（例如 RCA 反应的产物）物理结合，并形成可见的相互缠绕的聚集体，而在没有高分子量 NA（> 1 kb）的情况下没有观察到聚集体。肉眼检测到的目标 DNA 分子低至 0.62 amol (124 fM)。进一步应用于检测从 HeLa 细胞中分离的基因组 DNA 中的人乳头瘤病毒 18 型基因。这种检测方法灵敏且成本低，仪器最少，揭示了发展中国家和环境有限的地区分子诊断的巨大潜力。 (C) 2013 Elsevier B.V. 保留所有权利。
We are presenting a new method for sequence-specific and naked-eye detection of nucleic acids (NAs) through isothermal amplification and magnetic particle mediated aggregation. A padlock probe was designed to hybridize to specific sequence on target genes followed by ligation and rolling circle amplification (RCA). Magnetic particles (MPs) were then added into RCA products solution. MPs would physically combine with long linear DNA coils (such as products of RCA reaction) and form visible intertwined aggregates, while no aggregate was observed in the absence of high molecular weight NAs (> 1 kb). As little as 0.62 amol (124 fM) of target DNA molecules was detected by the naked eye. It was further applied to detect human papillomavirus type 18 gene in genomic DNA isolated from HeLa cells. This assay is sensitive and low-cost with minimal instrumentation, revealing great potential for molecular diagnostics in developing countries and regions with limited settings. (C) 2013 Elsevier B.V. All rights reserved.