Direct Detection of Conserved Viral Sequences and Other Nucleic Acid Motifs with Solid-State Nanopores.
Direct Detection of Conserved Viral Sequences and Other Nucleic Acid Motifs with Solid-State Nanopores.
复制标题
DOI：
10.1021/acsnano.0c10887
复制
发表时间：
2021-05-25
期刊：
影响因子：
17.1
通讯作者：
Hall AR
中科院分区：
文献类型：
--
作者：
Sethi K;Dailey GP;Zahid OK;Taylor EW;Ruzicka JA;Hall AR
快速可靠的核酸序列识别在基因分型、基因表达分析和病原体筛选等广泛领域中都是必不可少的。尤其是在病毒检测方面，这种能力对于最佳治疗反应和预防疾病传播至关重要。在这里，我们报告了一种基于固态纳米孔检测基因组规模单链DNA和RNA中识别序列基序的方法。通过设计与目标基因组中的目标序列具有互补性的DNA寡核苷酸探针，我们建立了一种协议，只有在目标存在的情况下，才能产生与探针相同长度的亲和标记双链分子。随后，该产物可以与蛋白质伴侣结合，并通过选择性的固态纳米孔分析进行定量分析。我们首先使用一个模型DNA基因组(M13mp18)来验证该方法，显示了成功分离和同时检测多个靶序列。然后，我们展示了通过识别和靶向人类免疫缺陷病毒(HIV-1B)中高度保守的序列来检测RNA病毒的协议。
Rapid and reliable recognition of nucleic acid sequences is essential to a broad range of fields including genotyping, gene expression analysis, and pathogen screening. For viral detection in particular, the capability is critical for optimal therapeutic response and preventing disease transmission. Here, we report an approach for detecting identifying sequence motifs within genome-scale single-strand DNA and RNA based on solid-state nanopores. By designing DNA oligonucleotide probes with complementarity to target sequences within a target genome, we establish a protocol to yield affinity-tagged duplex molecules the same length as the probe only if the target is present. The product can subsequently be bound to a protein chaperone and analyzed quantitatively with a selective solid-state nanopore assay. We first use a model DNA genome (M13mp18) to validate the approach, showing successful isolation and detection of multiple target sequences simultaneously. We then demonstrate the protocol for the detection of RNA viruses by identifying and targeting a highly conserved sequence within human immunodeficiency virus (HIV-1B).
登录
查看更多内容
影响因子：
14.9
作者：
Lassmann T;Frings O;Sonnhammer EL
通讯作者：
Sonnhammer EL
影响因子：
3.9
作者：
Anderson, Will;Lane, Rebecca;Trau, Matt
通讯作者：
Trau, Matt
影响因子：
56.9
作者：
FAUCI, AS
通讯作者：
FAUCI, AS
影响因子：
10.8
作者：
Carlsen, Autumn T.;Zahid, Osama K.;Hall, Adam R.
通讯作者：
Hall, Adam R.
影响因子：
3.7
作者：
Junqueira, Dennis Maletich;de Matos Almeida, Sabrina Esteves
通讯作者：
de Matos Almeida, Sabrina Esteves