Background

Hepatitis C Virus (HCV) is an asymptomatic chronic infection with serious clinical consequences. Timely and sensitive detection of HCV RNA is critical for infection control, treatment response follow up. While current technologies, such as PCR or isothermal amplification-based strategies, are specific, they are expensive, labor intensive, time consuming limiting their use in field and small laboratories.

Results

We introduce a novel technology for detecting nucleic acids, as exemplified by HCV RNA, in clinical specimens. This approach utilizes the crosslinked Enhanced Emission (CEE) phenomena upon mixing fluorescent amino functionalized silica-coated nitrogen-doped carbon dots (N-CDs/SiO₂/NH₂) with magnetic extracted unamplified HCV RNA showed a significant and immediate fluorescence enhancement. This method was integrated into a semi-automated 3D printed microfluidic chip, wherein the unamplified RNA was mixed with the N-CDs/SiO₂/NH₂. The assay was also employed on the conventional 96-well plate format. This assay offers high sensitivity with detection limit of 500 IU/ml and 1000 IU/ml for the chip and well plate respectively. The sample-to-result time was < 20 minutes on the chip and is simpler than the amplification-based techniques. Analyzing 141 patient samples yielded sensitivity and specificity of 96.47 % and 98.79 % respectively.

Significance

This application of N-CDs/SiO2 as nucleic acid fluorescent probes for the first time, offers a versatile, cost-effective, and universal platform for nucleic acids detection. The developed system can be employed in conventional microwell plate-based detection with minimal modifications to current laboratory setups. Additionally, it was integrated into a 3D-printed microfluidic chip enabling enhancement in specificity, sensitivity and accuracy.

中文翻译：

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基于微流控的硅胶包埋碳点荧光增强，用于直接检测和定量临床样品中未扩增的 HCV RNA

 背景

丙型肝炎病毒 （HCV） 是一种无症状的慢性感染，具有严重的临床后果。及时灵敏地检测 HCV RNA 对于感染控制和治疗反应随访至关重要。虽然当前的技术（如 PCR 或基于等温扩增的策略）是特定的，但它们价格昂贵、劳动密集、耗时，限制了它们在现场和小型实验室中的使用。

 结果

我们介绍了一种检测临床标本中核酸的新技术，以 HCV RNA 为例。该方法利用交联增强发射 （CEE） 现象，将荧光氨基官能化二氧化硅包被的氮掺杂碳点 （N-CDs/SiO₂/NH₂） 与磁性提取的未扩增 HCV RNA 混合后显示出显着且即时的荧光增强。该方法被集成到半自动 3D 打印微流控芯片中，其中未扩增的 RNA 与 N-CDs/SiO₂/NH₂ 混合。该分析也用于常规的 96 孔板格式。该测定具有高灵敏度，芯片和孔板的检测限分别为 500 IU/ml 和 1000 IU/ml。在芯片上，样品到结果的时间为 < 20 分钟，比基于扩增的技术更简单。分析 141 份患者样本的敏感性和特异性分别为 96.47 % 和 98.79 %。

 意义

N-CDs/SiO2 首次用作核酸荧光探针，为核酸检测提供了一个多功能、经济高效且通用的平台。开发的系统可用于传统的基于微孔板的检测，只需对当前的实验室设置进行最少的修改。此外，它还被集成到 3D 打印的微流体芯片中，从而提高了特异性、灵敏度和准确性。