CAS Engineering Laboratory for Nanozyme, Institute of Biophysics, Chinese Academic of Science, Beijing, 100101, China.
CAS Engineering Laboratory for Nanozyme, Institute of Biophysics, Chinese Academic of Science, Beijing, 100101, China; School of Basic Medical Science, Southwest Medical University, Luzhou, 646000, China.
Biosens Bioelectron. 2022 Dec 1;217:114739. doi: 10.1016/j.bios.2022.114739. Epub 2022 Sep 19.
The coronavirus disease 2019 (COVID-19) pandemic has created a huge demand for sensitive and rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The current gold standard for SARS-CoV-2 detection is reverse transcription-polymerase chain reaction (RT-PCR)-based nucleic acid amplification. However, RT-PCR is time consuming and requires specialists and large instruments that are unattainable for point-of-care testing (POCT). To develop POCT for SARS-CoV-2, we combined recombinase polymerase amplification (RPA) and FeS nanozyme strips to achieve facile nucleic acid amplification and subsequent colorimetric signal enhancement based on the high peroxidase-like activity of the FeS nanozymes. This method showed a nucleic acid limit of detection (LOD) for SARS-CoV-2 of 200 copies/mL, close to that of RT-PCR. The unique catalytic properties of the FeS nanozymes enabled the nanozyme-strip to amplify colorimetric signals via the nontoxic 3,3',5,5'-tetramethylbenzidine (TMB) substrate. Importantly, the detection of clinical samples of human papilloma virus type 16 (HPV-16) showed 100% agreement with previous RT-PCR results, highlighting the versatility and reliability of this method. Our findings suggest that nanozyme-based nucleic acid detection has great potential in the development of POCT diagnosis for COVID-19 and other viral infections.
2019 年冠状病毒病(COVID-19)大流行对严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)的敏感和快速检测提出了巨大需求。目前 SARS-CoV-2 检测的金标准是基于逆转录-聚合酶链反应(RT-PCR)的核酸扩增。然而,RT-PCR 耗时且需要专家和大型仪器,而这些在即时检测(POCT)中是无法实现的。为了开发 SARS-CoV-2 的 POCT,我们将重组酶聚合扩增(RPA)和 FeS 纳米酶片结合在一起,实现了简便的核酸扩增,随后基于 FeS 纳米酶的高过氧化物酶样活性进行了比色信号增强。该方法对 SARS-CoV-2 的核酸检测限(LOD)为 200 拷贝/mL,接近 RT-PCR 的水平。FeS 纳米酶的独特催化特性使纳米酶片能够通过非毒性 3,3',5,5'-四甲基联苯胺(TMB)底物扩增比色信号。重要的是,对人乳头瘤病毒 16 型(HPV-16)临床样本的检测与之前的 RT-PCR 结果完全一致,突出了该方法的多功能性和可靠性。我们的研究结果表明,基于纳米酶的核酸检测在 COVID-19 和其他病毒感染的 POCT 诊断开发方面具有巨大潜力。
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