Institute of Marine Science and Technology, Shandong University, Qingdao, Shandong, 266237, P.R. China.
Chemistry. 2022 Mar 28;28(18):e202104054. doi: 10.1002/chem.202104054. Epub 2022 Mar 8.
The ongoing outbreak of the severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) has spread globally and poses a threat to public health and National economic development. Rapid and high-throughput SARS-CoV-2 RNA detection without the need of RNA extraction and amplification remain a key challenge. In this study, a new SARS-CoV-2 RNA detection strategy using a microfluidic biochip for the rapid and ultrasensitive detection of SARS-CoV-2 without RNA extraction and amplification was developed. This new strategy takes advantage of the specific SARS-CoV-2 RNA and probe DNA reaction in the microfluidic channel, fluorescence signal regulation by nanomaterials, and accurate sample control by the microfluidic chip. It presents an ultralow limit of detection of 600 copies mL in a large linear detection regime from 1 aM to 100 fM. Fifteen samples were simultaneously detected in 40 min without the need for RNA purification and amplification. The detection accuracy of the strategy was validated through quantitative reverse transcription polymerase chain reaction (qRT-PCR), with a recovery of 99-113 %. Therefore, the SARS-CoV-2 RNA detection strategy proposed in this study can potentially be used for the quantitative diagnosis of viral infectious diseases.
持续爆发的严重急性呼吸综合征冠状病毒 2（SARS-CoV-2）已在全球范围内传播，对公众健康和国家经济发展构成威胁。无需 RNA 提取和扩增即可快速、高通量检测 SARS-CoV-2 RNA 仍然是一个关键挑战。在这项研究中，开发了一种新的 SARS-CoV-2 RNA 检测策略，该策略使用微流控生物芯片在无需 RNA 提取和扩增的情况下快速、超灵敏地检测 SARS-CoV-2。这种新策略利用了微流道中 SARS-CoV-2 RNA 和探针 DNA 的特异性反应、纳米材料对荧光信号的调控以及微流控芯片对样品的精确控制。它在 1 aM 至 100 fM 的大线性检测范围内呈现出 600 拷贝/ml 的超低检测限。无需 RNA 纯化和扩增，即可在 40 分钟内同时检测 15 个样本。该策略的检测准确性通过定量逆转录聚合酶链反应（qRT-PCR）进行了验证，回收率为 99-113%。因此，本研究提出的 SARS-CoV-2 RNA 检测策略有望用于病毒感染性疾病的定量诊断。
