Department of Biochemistry, Faculty of Science, Ege University, 35100, Bornova, Izmir, Turkey.
Central Research Test and Analysis Laboratory Application and Research Center, Ege University, 35100, Bornova, Izmir, Turkey.
Talanta. 2022 Jun 1;243:123356. doi: 10.1016/j.talanta.2022.123356. Epub 2022 Mar 3.
The increasing mutation frequency of the SARS-CoV-2 virus and the emergence of successive variants have made correct diagnosis hard to perform. Developing efficient and accurate methods to diagnose infected patients is crucial to effectively mitigate the pandemic. Here, we developed an electrochemical immunosensor based on SARS-CoV-2 antibody cocktail-conjugated magnetic nanoparticles for the sensitive and accurate detection of the SARS-CoV-2 virus and its variants in nasopharyngeal swabs. The application of the antibody cocktail was compared with commercially available anti-SARS-CoV-2 S1 (anti-S1) and anti-S2 monoclonal antibodies. After optimization and calibration, the limit of detection (LOD) determination demonstrated a LOD = 0.53-0.75 ng/mL for the antibody cocktail-based sensor compared with 0.93 ng/mL and 0.99 ng/mL for the platforms using anti-S1 and anti-S2, respectively. The platforms were tested with human nasopharyngeal swab samples pre-diagnosed with RT-PCR (10 negatives and 40 positive samples). The positive samples include the original, alpha, beta, and delta variants (n = 10, for each). The polyclonal antibody cocktail performed better than commercial anti-S1 and anti-S2 antibodies for all samples reaching 100% overall sensitivity, specificity, and accuracy. It also showed a wide range of variants detection compared to monoclonal antibody-based platforms. The present work proposes a versatile electrochemical biosensor for the indiscriminate detection of the different variants of SARS-CoV-2 using a polyclonal antibody cocktail. Such diagnostic tools allowing the detection of variants can be of great efficiency and economic value in the fight against the ever-changing SARS-CoV-2 virus.
新冠病毒(SARS-CoV-2)的突变频率不断增加,连续出现的变异株使得准确诊断变得困难。开发高效、准确的方法来诊断感染患者对于有效缓解大流行至关重要。在这里,我们开发了一种基于 SARS-CoV-2 抗体鸡尾酒偶联磁纳米粒子的电化学免疫传感器,用于灵敏、准确地检测鼻咽拭子中的 SARS-CoV-2 病毒及其变异株。我们将抗体鸡尾酒的应用与市售的抗 SARS-CoV-2 S1(抗 S1)和抗 S2 单克隆抗体进行了比较。经过优化和校准,基于抗体鸡尾酒的传感器的检测限(LOD)测定结果表明,与使用抗 S1 和抗 S2 的平台相比,LOD 分别为 0.53-0.75ng/mL 和 0.93ng/mL。我们用经 RT-PCR 预诊断的人类鼻咽拭子样本(10 个阴性和 40 个阳性样本)对这些平台进行了测试。阳性样本包括原始、阿尔法、贝塔和德尔塔变异株(n=10,每个)。与商业的抗 S1 和抗 S2 抗体相比,多克隆抗体鸡尾酒在所有样本中的性能都更好,总体敏感性、特异性和准确性均达到 100%。与基于单克隆抗体的平台相比,它还显示出更广泛的变异株检测范围。本工作提出了一种通用的电化学生物传感器,使用多克隆抗体鸡尾酒可对 SARS-CoV-2 的不同变异株进行无差别检测。这种能够检测变异株的诊断工具在对抗不断变化的 SARS-CoV-2 病毒方面具有巨大的效率和经济价值。