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利用血清型鉴定能力检测登革热病毒 DNA 的飞摩尔水平

Femtomolar Detection of Dengue Virus DNA with Serotype Identification Ability.

机构信息

Laboratory of Biotechnology, Research Institute of Green Science and Technology , Shizuoka University , 836 Ohya , Suruga-ku, Shizuoka 422-8529 , Japan.

Department of Biomedical Engineering and Environmental Sciences , National Tsing Hua University , 101 Section 2, Kuang-Fu Road , Hsinchu , 30013 , Taiwan.

出版信息

Anal Chem. 2018 Nov 6;90(21):12464-12474. doi: 10.1021/acs.analchem.8b01802. Epub 2018 Oct 12.

Abstract

Dengue surveillance trusts only on reverse transcription-polymerase chain reaction (RT-PCR) type methodologies for confirmation of dengue virus serotypes; however, its real time application is restricted due to the expensive, complicated, and time-consuming process. In search of a new sensing system, here, we have reported a two-way-detection method for Dengue virus (DENV) serotype identification along with DNA quantification by using a new class of nanocomposite of gold nanoparticles (AuNP) and nitrogen, sulfur codoped graphene quantum dots (N,S-GQDs). The N,S-GQDs@AuNP has been used for serotype detection via a simple fluorescence technique using four dye-combined probe DNAs which is further validated by confocal microscopy. The quantification of the DNA has been measured by the differential pulse voltammetric (DPV) technique using methyelene blue as a redox indicator. Results obtained in this study, clearly demonstrate that the N,S-GQDs@AuNP can efficiently detect the four serotypes of DENV individually in the concentration range of 10 to 10 M with the LOD of 9.4 fM. In addition, to confirm its applicability in long chained complex DNA system, the sensor was also applied to the clinically isolated DENV DNA and showed satisfactory performances for serotype identification as well as quantification. We hope this simple and reliable method can pave an avenue for the development of sensitive and robust sensing probes in biomedical applications.

摘要

登革热监测仅依赖于逆转录-聚合酶链反应 (RT-PCR) 方法来确认登革病毒血清型;然而,由于其昂贵、复杂和耗时的过程,其实时应用受到限制。在寻找新的传感系统的过程中,在这里,我们报告了一种用于登革病毒 (DENV) 血清型鉴定以及 DNA 定量的双向检测方法,该方法使用了一种新型的金纳米粒子 (AuNP) 和氮、硫共掺杂石墨烯量子点 (N,S-GQDs) 的纳米复合材料。通过使用四种染料组合探针 DNA 的简单荧光技术,可以对 N,S-GQDs@AuNP 进行血清型检测,并用共聚焦显微镜进行了进一步验证。使用亚甲基蓝作为氧化还原指示剂,通过差分脉冲伏安法 (DPV) 技术测量 DNA 的定量。本研究的结果清楚地表明,N,S-GQDs@AuNP 可以在 10 到 10 M 的浓度范围内分别有效地检测出四种 DENV 血清型,LOD 为 9.4 fM。此外,为了确认其在长链复杂 DNA 系统中的适用性,该传感器还应用于临床分离的 DENV DNA,并且在血清型鉴定和定量方面表现出令人满意的性能。我们希望这种简单可靠的方法可以为生物医学应用中灵敏和强大的传感探针的发展铺平道路。

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