State Key Laboratory of NBC Protection for Civilian, Beijing 102205, China.
Sensors (Basel). 2022 Mar 21;22(6):2402. doi: 10.3390/s22062402.
To satisfy the need to develop highly sensitive methods for detecting the severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) and further enhance detection efficiency and capability, a new method was created for detecting SARS-CoV-2 of the open reading frames 1ab (ORF1ab) target gene by a electrochemiluminescence (ECL) biosensor based on dual-probe hybridization through the use of a detection model of "magnetic capture probes-targeted nucleic acids-Ru(bpy) labeled signal probes". The detection model used magnetic particles coupled with a biotin-labeled complementary nucleic acid sequence of the SARS-CoV-2 ORF1ab target gene as the magnetic capture probes and Ru(bpy) labeled amino modified another complementary nucleic acid sequence as the signal probes, which combined the advantages of the highly specific dual-probe hybridization and highly sensitive ECL biosensor technology. In the range of 0.1 fM~10 µM, the method made possible rapid and sensitive detection of the ORF1ab gene of the SARS-CoV-2 within 30 min, and the limit of detection (LOD) was 0.1 fM. The method can also meet the analytical requirements for simulated samples such as saliva and urine with the definite advantages of a simple operation without nucleic acid amplification, high sensitivity, reasonable reproducibility, and anti-interference solid abilities, expounding a new way for efficient and sensitive detection of SARS-CoV-2.
为满足开发高灵敏度方法检测严重急性呼吸综合征冠状病毒 2 型（SARS-CoV-2）的需求，并进一步提高检测效率和能力，创建了一种基于双探针杂交的电化学发光（ECL）生物传感器检测 SARS-CoV-2 的开放阅读框 1ab（ORF1ab）靶基因的新方法，检测模型为“磁捕获探针-靶向核酸-Ru（bpy）标记信号探针”。检测模型使用与 SARS-CoV-2 ORF1ab 靶基因的生物素标记互补核酸序列偶联的磁性颗粒作为磁捕获探针，Ru（bpy）标记的氨基修饰另一条互补核酸序列作为信号探针，结合了高度特异的双探针杂交和高灵敏度的 ECL 生物传感器技术的优势。在 0.1 fM~10 µM 的范围内，该方法可在 30 分钟内快速灵敏地检测 SARS-CoV-2 的 ORF1ab 基因，检测限（LOD）为 0.1 fM。该方法还可以满足唾液和尿液等模拟样本的分析要求，具有操作简单、无需核酸扩增、高灵敏度、合理重现性和抗干扰固体能力等明显优势，为 SARS-CoV-2 的高效灵敏检测开辟了新途径。
