Gupta Ankit, Anand Anjali, Jain Neha, Goswami Sandeep, Anantharaj Anbalagan, Patil Sharanabasava, Singh Rahul, Kumar Amit, Shrivastava Tripti, Bhatnagar Shinjini, Medigeshi Guruprasad R, Sharma Tarun Kumar
Aptamer Technology and Diagnostics Laboratory, Multidisciplinary Clinical and Translational Research, Translational Health Science and Technology Institute, Faridabad, Haryana 121001, India.
Discipline of Biosciences and Biomedical Engineering, Indian Institute of Technology Indore, Simrol, Madhya Pradesh 453552, India.
Mol Ther Nucleic Acids. 2021 Jun 24;26:321-332. doi: 10.1016/j.omtn.2021.06.014. eCollection 2021 Dec 3.
The recent SARS-CoV-2 outbreak has been declared a global health emergency. It will take years to vaccinate the whole population to protect them from this deadly virus, hence the management of SARS-CoV-2 largely depends on the widespread availability of an accurate diagnostic test. Toward addressing the unmet need of a reliable diagnostic test in the current work by utilizing the power of Systematic Evolution of Ligands by EXponential enrichment, a 44-mer G-quadruplex-forming DNA aptamer against spike trimer antigen of SARS-CoV-2 was identified. The lead aptamer candidate (S14) was characterized thoroughly for its binding, selectivity, affinity, structure, and batch-to-batch variability by utilizing various biochemical, biophysical, and techniques. S14 has demonstrated a low nanomolar K, confirming its tight binding to a spike antigen of SARS-CoV-2. S14 can detect as low as 2 nM of antigen. The clinical evaluation of S14 aptamer on nasopharyngeal swab specimens (n = 232) has displayed a highly discriminatory response between SARS-CoV-2 infected individuals from the non-infected one with a sensitivity and specificity of ∼91% and 98%, respectively. Importantly, S14 aptamer-based test has evinced a comparable performance with that of RT-PCR-based assay. Altogether, this study established the utility of aptamer technology for the detection of SARS-CoV-2.
最近爆发的新型冠状病毒肺炎已被宣布为全球卫生紧急事件。要为全体人口接种疫苗以保护他们免受这种致命病毒的侵害需要数年时间，因此新型冠状病毒肺炎的防控很大程度上依赖于广泛可得的准确诊断检测方法。为了通过指数富集的配体系统进化技术满足当前工作中对可靠诊断检测方法的未满足需求，我们鉴定出了一种针对新型冠状病毒刺突三聚体抗原的44聚体G-四链体形成DNA适配体。通过各种生化、生物物理技术，对主要的适配体候选物（S14）的结合、选择性、亲和力、结构和批次间变异性进行了全面表征。S14的解离常数低至纳摩尔级别，证实了它与新型冠状病毒刺突抗原的紧密结合。S14能够检测低至2 nM的抗原。对232份鼻咽拭子标本进行的S14适配体临床评估显示，在新型冠状病毒感染个体与未感染个体之间呈现出高度区分性反应，敏感性和特异性分别约为91%和98%。重要的是，基于S14适配体的检测方法表现出与基于逆转录聚合酶链反应的检测方法相当的性能。总之，本研究确立了适配体技术在新型冠状病毒检测中的实用性。
