基于纳米孔技术检测唾液样本中的牙周炎生物标志物 miR31。

IF 3 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS

ELECTROPHORESIS Pub Date : 2024-08-21 DOI:10.1002/elps.202400134

Pearl Arora, Haiyan Zheng, Sathishkumar Munusamy, Rana Jahani, Xiyun Guan

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引用次数: 0

摘要

微RNA（miRNA）在转录后基因调控中发挥着重要作用。miRNA 水平的异常是导致包括牙周炎在内的各种疾病的原因。因此，灵敏、特异、准确地检测与疾病相关的 miRNA 对早期诊断至关重要，同时也有助于抑制剂筛选和药物设计。在这项研究中，我们开发了一种无标记的实时传感方法，利用工程蛋白纳米孔，在互补 ssDNA 作为分子探针的情况下，检测经常与牙周炎相关的 miR31。我们的方法快速且灵敏度高，可在几分钟内检测出纳摩尔浓度的 miR31。此外，即使存在干扰核酸，我们的传感器对目标 miR31 序列也有很高的选择性。此外，我们还成功分析了人工唾液和人类唾液样本。我们开发的纳米孔传感平台可用于检测其他 miRNA，并有望应用于疾病生物标志物的临床诊断。

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Nanopore-based detection of periodontitis biomarker miR31 in saliva samples.

MicroRNAs (miRNAs) play important roles in posttranscriptional gene regulation. Aberrations in the miRNA levels have been the cause behind various diseases, including periodontitis. Therefore, sensitive, specific, and accurate detection of disease-associated miRNAs is vital to early diagnosis and can facilitate inhibitor screening and drug design. In this study, we developed a label-free, real-time sensing method for the detection of miR31, which has been frequently linked to periodontitis, using an engineered protein nanopore and in the presence of a complementary ssDNA as a molecular probe. Our method is rapid and highly sensitive with nanomolar concentration of miR31 that could be determined in minutes. Furthermore, our sensor showed high selectivity toward the target miR31 sequence even in the presence of interfering nucleic acids. In addition, artificial saliva and human saliva samples were successfully analyzed. Our developed nanopore sensing platform could be used to detect other miRNAs and offers a potential application for the clinical diagnosis of disease biomarkers.

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来源期刊

ELECTROPHORESIS 生物-分析化学

CiteScore

6.30

自引率

13.80%

发文量

244

审稿时长

1.9 months

期刊介绍： ELECTROPHORESIS is an international journal that publishes original manuscripts on all aspects of electrophoresis, and liquid phase separations (e.g., HPLC, micro- and nano-LC, UHPLC, micro- and nano-fluidics, liquid-phase micro-extractions, etc.). Topics include new or improved analytical and preparative methods, sample preparation, development of theory, and innovative applications of electrophoretic and liquid phase separations methods in the study of nucleic acids, proteins, carbohydrates natural products, pharmaceuticals, food analysis, environmental species and other compounds of importance to the life sciences. Papers in the areas of microfluidics and proteomics, which are not limited to electrophoresis-based methods, will also be accepted for publication. Contributions focused on hyphenated and omics techniques are also of interest. Proteomics is within the scope, if related to its fundamentals and new technical approaches. Proteomics applications are only considered in particular cases. Papers describing the application of standard electrophoretic methods will not be considered. Papers on nanoanalysis intended for publication in ELECTROPHORESIS should focus on one or more of the following topics: • Nanoscale electrokinetics and phenomena related to electric double layer and/or confinement in nano-sized geometry • Single cell and subcellular analysis • Nanosensors and ultrasensitive detection aspects (e.g., involving quantum dots, "nanoelectrodes" or nanospray MS) • Nanoscale/nanopore DNA sequencing (next generation sequencing) • Micro- and nanoscale sample preparation • Nanoparticles and cells analyses by dielectrophoresis • Separation-based analysis using nanoparticles, nanotubes and nanowires.