Anantharaj Anbalagan, Das Soon Jyoti, Sharanabasava Patil, Lodha Rakesh, Kabra Sushil K, Sharma Tarun Kumar, Medigeshi Guruprasad R
National Capital Region - Biotech Science Cluster, Translational Health Science and Technology Institute (THSTI), Faridabad, India.
Department of Pediatrics, All India Institute of Medical Sciences, New Delhi, India.
Front Mol Biosci. 2020 Dec 23;7:586254. doi: 10.3389/fmolb.2020.586254. eCollection 2020.
The gold standard for the diagnosis of SARS-CoV-2, the causative agent of COVID-19, is real-time polymerase chain reaction (PCR), which is labor-intensive, expensive, and not widely available in resource-poor settings. Therefore, it is imperative to develop novel, accurate, affordable, and easily accessible assays/sensors to diagnose and isolate COVID-19 cases. To address this unmet need, we utilized the catalytic potential of peroxidase-like DNAzyme and developed a simple visual detection assay for SARS-CoV-2 RNA using a conventional thermal cycler by the PCR-induced generation of DNAzyme sensor. The performance of RT-PCR DNAzyme-based sensor was comparable to that of real-time PCR. The pilot scale validation of RT-PCR DNAzyme-based sensor has shown ~100% sensitivity and specificity in clinical specimens (nasopharyngeal swab, = 34), with a good correlation (Spearman = 0.799) with the Ct-value of fluorescence probe-based real-time PCR. These findings clearly indicate the potential of this inexpensive, sensitive, and specific molecular diagnostic test to extend our testing capabilities for the detection of SARS-CoV-2 to curtail COVID-19 transmission.
新冠病毒病（COVID-19）的病原体严重急性呼吸综合征冠状病毒2（SARS-CoV-2）诊断的金标准是实时聚合酶链反应（PCR），该方法劳动强度大、成本高，且在资源匮乏地区无法广泛使用。因此，开发新颖、准确、经济且易于使用的检测方法/传感器用于诊断和隔离COVID-19病例势在必行。为满足这一未被满足的需求，我们利用了类过氧化物酶DNAzyme的催化潜力，通过PCR诱导产生DNAzyme传感器，使用传统热循环仪开发了一种用于SARS-CoV-2 RNA的简单可视化检测方法。基于RT-PCR DNAzyme的传感器性能与实时PCR相当。基于RT-PCR DNAzyme的传感器的中试规模验证在临床标本（鼻咽拭子，n = 34）中显示出约100%的灵敏度和特异性，与基于荧光探针的实时PCR的Ct值具有良好的相关性（Spearman相关系数 = 0.799）。这些发现清楚地表明了这种廉价、灵敏且特异的分子诊断测试在扩展我们检测SARS-CoV-2以遏制COVID-19传播的检测能力方面的潜力。
