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用于寨卡病毒荧光分子诊断的本征导电聚合物混合双层膜。

Intrinsically conductive polymers hybrid bilayer films for the fluorescence molecular diagnosis of the Zika virus.

机构信息

Pós-Graduação em Ciência de Materiais, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.

Departamento de Física, Universidade Federal de Pernambuco, 50670-901 Recife, PE, Brazil.

出版信息

Colloids Surf B Biointerfaces. 2021 Dec;208:112120. doi: 10.1016/j.colsurfb.2021.112120. Epub 2021 Sep 16.

Abstract

In 2016, the Zika virus (ZIKV) infection became a major public health problem, after the discovery that an alarming increase in the number of Brazilian newborns with microcephaly could be associated with the occurrence of this viral disease during the pregnancy of their mothers. The urgent need for simple diagnostic methods that allow rapid screening of suspected cases has stimulated the search for low-cost devices capable of detecting specific sequences of nucleic acids. The present work describes the development of nanostructured films formed by bilayers of conjugated polymers for rapid detection of the presence of Zika virus DNA, via fluorescence methods. For this, we initially deposited alternating layers of polyaniline (PANI) and polypyrrole (PPY) on the surface of polyethylene terephthalate (PET) sheets. The films obtained were then characterized by SEM, UV-Vis, ATR-FTIR, and contact angle measurements. For their use as quenchers for the diagnosis of Zika, a single DNA strand-specific for ZIKV was labeled with a fluorophore (FAM-ssDNA). We determined the time required for the saturation of the interaction between probe FAM-ssDNA and the film (180 min) and the time for the maximal hybridization between FAM-ssDNA and target DNA to occur (60 min). The detection limits were estimated as 345 pM and 278 pM for the PET/PPY-PANI and PET/PANI-PPY hybrid films, respectively. The simplicity of the procedure, coupled with the fact that a positive/negative response can be obtained in less than 60 min, suggests that the proposal of using these polymeric bilayer films is a promising methodology for the development of rapid molecular diagnostic tests.

摘要

2016 年,寨卡病毒(ZIKV)感染成为一个主要的公共卫生问题,此前发现巴西新生儿小头畸形数量的惊人增加可能与母亲怀孕期间发生这种病毒病有关。对能够快速筛选疑似病例的简单诊断方法的迫切需求,刺激了对能够检测特定核酸序列的低成本设备的寻找。本工作描述了通过荧光方法快速检测寨卡病毒 DNA 存在的由共轭聚合物双层形成的纳米结构薄膜的开发。为此,我们首先在聚对苯二甲酸乙二醇酯(PET)片材的表面上沉积交替的聚苯胺(PANI)和聚吡咯(PPY)层。然后通过 SEM、UV-Vis、ATR-FTIR 和接触角测量对获得的薄膜进行了表征。为了将其用作寨卡诊断的猝灭剂,用荧光团(FAM-ssDNA)标记了针对 ZIKV 的单链 DNA 探针。我们确定了探针 FAM-ssDNA 与薄膜之间的相互作用达到饱和所需的时间(180 分钟)以及 FAM-ssDNA 与靶 DNA 之间发生最大杂交所需的时间(60 分钟)。对于 PET/PPY-PANI 和 PET/PANI-PPY 杂化薄膜,检测限分别估计为 345 pM 和 278 pM。该过程的简单性,加上可以在不到 60 分钟内获得阳性/阴性反应,表明使用这些聚合物双层薄膜的建议是开发快速分子诊断测试的一种很有前途的方法。

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