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基于铂纳米颗粒(PtNPs)的 CRISPR/Cas12a 平台,用于检测临床样本中的核酸和蛋白质。

Platinum nanoparticles (PtNPs)-based CRISPR/Cas12a platform for detection of nucleic acid and protein in clinical samples.

机构信息

Guangdong Province Engineering Research Center of Antibody Drug and Immunoassay, Department of Bioengineering, College of Life Science and Technology, Jinan University, Guangzhou, 510632, China; Department of Oncology, The First Affiliated Hospital, Jinan University, Guangzhou, 510632, China; Guangdong Biowings Tech Limited, Foshan, 528000, China.

Guangdong Province Engineering Research Center of Antibody Drug and Immunoassay, Department of Bioengineering, College of Life Science and Technology, Jinan University, Guangzhou, 510632, China.

出版信息

Anal Chim Acta. 2022 Sep 8;1225:340203. doi: 10.1016/j.aca.2022.340203. Epub 2022 Aug 11.

Abstract

Early rapid screening diagnostic assay is essential for the identification, prevention, and evaluation of many contagious or refractory diseases. The optical density transducer created by platinum nanoparticles (PtNPs) (OD-CRISPR) is reported in the present research as a cheap and easy-to-execute CRISPR/Cas12a-based diagnostic platform. The OD-CRISPR uses PtNPs, with ultra-high peroxidase-mimicking activity, to increase the detection sensitivity, thereby enabling the reduction of detection time and cost. The OD-CRISPR can be utilized to identify nucleic acid or protein biomarkers within an incubation time of 30-40min in clinical specimens. In the case of taking severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) N gene as an instance, when compared to a quantitative reverse transcription-polymerase chain reaction (RT-qPCR), the OD-CRISPR test attains a sensitivity of 79.17% and a specificity of 100%. In terms of detecting prostate-specific antigen (PSA), aptamer-based OD-CRISPR assay achieves the least discoverable concentration of 0.01 ng mL. In general, the OD-CRISPR can detect nucleic acid and protein biomarkers, and is a potential strategy for early rapid screening diagnostic tools.

摘要

早期快速筛选诊断检测对于识别、预防和评估许多传染性或难治性疾病至关重要。本研究报道了一种由铂纳米粒子(PtNPs)(OD-CRISPR)制成的光密度传感器,它是一种廉价且易于执行的基于 CRISPR/Cas12a 的诊断平台。OD-CRISPR 利用具有超高过氧化物酶模拟活性的 PtNPs 提高了检测灵敏度,从而能够缩短检测时间和降低成本。OD-CRISPR 可以在 30-40 分钟的孵育时间内用于识别临床标本中的核酸或蛋白质生物标志物。以严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)N 基因为例,与定量逆转录聚合酶链反应(RT-qPCR)相比,OD-CRISPR 测试的灵敏度为 79.17%,特异性为 100%。在检测前列腺特异性抗原(PSA)方面,基于适体的 OD-CRISPR 检测法达到了 0.01ng/mL 的最低检测浓度。总的来说,OD-CRISPR 可以检测核酸和蛋白质生物标志物,是一种用于早期快速筛选诊断工具的潜在策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f918/9365833/c56e90f71120/ga1_lrg.jpg

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