Rapid detection of SARS-CoV-2 using a radiolabeled antibody.
Rapid detection of SARS-CoV-2 using a radiolabeled antibody.
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DOI：
10.1016/j.nucmedbio.2021.05.002
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发表时间：
2021-07
影响因子：
3.1
通讯作者：
Pillarsetty NVK
中科院分区：
文献类型：
--
作者：
Pirovano G;Ordonez AA;Jain SK;Reiner T;Carroll LS;Pillarsetty NVK
严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）感染是2019冠状病毒病（COVID-19）的病因，对人类构成严重风险，并对全球医疗保健系统构成巨大挑战。自COVID-19大流行初期以来，充分的检测显然是限制和控制SARS-CoV-2传播的重要步骤。在这里，我们提出了一个准确，廉价，可扩展，便携式，快速检测试剂盒，直接检测SARS-CoV-2的生物样本，甚至可以转化为人口测试。我们已经证明，我们的方法可以可靠地识别病毒载量，并可用于接触那些无法获得更复杂和昂贵测试的人群。拟议的SARS-CoV-2检测试剂盒基于SARS-CoV-2靶向抗体（CR 3022）的组合，该抗体靶向病毒表面的刺突蛋白S1结构域。这种抗体用长寿命同位素（碘-125）进行放射性标记，使我们能够检测SARS-CoV-2样本中的结合抗体。我们使用了一系列体外试验来确定灵敏度和特异性，并促进检测试剂盒的自动化。通过离心步骤和半透膜从唾液样品中提取结合的抗体。我们的试剂盒使用SARS-CoV-2病毒粒子进一步验证。我们能够可靠地完成[125 I] I-CR 3022的放射合成而不损失结合。SARS-CoV-2-sensing抗体显示出保持其刺突S1亲和力，并结合低至2.5-5 ng的刺突蛋白。然后，我们使用珠结合钉S1开发的分离试剂盒，这被证明是易于使用和廉价的。该试剂盒使得从唾液样样品中提取结合抗体成为可能。我们能够使用完整的SARS-CoV-2病毒粒子验证分离试剂盒，并表明我们的试剂盒可以检测低至19，700 PFU/mL（~ 9.22%TBF）和高达1，970，000 PFU/mL（45.04%TBF）的病毒浓度。在这里，我们报告的开发和验证的SARS-CoV-2的检测系统的基础上结合的特异性放射性标记的抗体和分离膜。我们证明了我们的系统与FDA已经批准的其他SARS-CoV-2检测试剂盒相当，并相信这项技术可以很容易地部署到资源有限的国家用于诊断COVID-19。此外，工作流程可以很容易地适应于靶向其他抗原，从而靶向其他类型的疾病。
Infection by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of coronavirus 2019 disease (COVID-19), poses a serious risk to humanity and represents a huge challenge for healthcare systems worldwide. Since the early days of the COVID-19 pandemic, it has been evident that adequate testing is an essential step in limiting and controlling the spread of SARS-CoV-2. Here, we present an accurate, inexpensive, scalable, portable, and rapid detection kit to directly detect SARS-CoV-2 in biological samples that could even be translated for population testing. We have demonstrated that our method can reliably identify viral load and could be used to reach those fractions of the population with limited access to more sophisticated and expensive tests. The proposed SARS-CoV-2 detection kit is based on the combination of a SARS-CoV-2-targeted antibody (CR3022) that targets spike protein S1 domain on the viral surface. This antibody was radiolabeled with a long-lived isotope (Iodine-125) to allow us to detect bound antibody in samples with SARS-CoV-2. We used a series of in vitro assays to determine sensitivity and specificity and facilitate automation of the testing kit. Bound antibody was extracted from saliva samples via a centrifugation step and a semi-permeable membrane. Our kit was further validated using SARS-CoV-2 virions. We were able to accomplish radiosynthesis of [125I]I-CR3022 reliably without loss of binding. The SARS-CoV-2-sensing antibody was shown to maintain its spike S1 affinity and to bind to as low as 2.5–5 ng of spike protein. We then used beads-bound spike S1 to develop a separation kit which proved to be both easy to use and inexpensive. The kit made it possible to extract bound antibody from the saliva-like sample. We were able to validate the separation kit using intact SARS-CoV-2 virions and showed that our kit can detect a viral concentration as low as 19,700 PFU/mL (~ 9.22%TBF) and as high as 1,970,000 PFU/mL (45.04%TBF). Here we report the development and validation of a SARS-CoV-2 detection system based on the combination of a specific radiolabeled antibody and a separation membrane. We demonstrate our system to be comparable to other SARS-CoV-2 detection kits already approved by the FDA and believe this technology could be easily deployed to countries with limited resources for the diagnosis of COVID-19. Furthermore, workflows could be easily adapted to target other antigens and therefore other types of diseases.
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影响因子：
16.6
作者：
Willem L;Abrams S;Libin PJK;Coletti P;Kuylen E;Petrof O;Møgelmose S;Wambua J;Herzog SA;Faes C;Beutels P;Hens N
通讯作者：
Hens N
DOI：
10.1038/s41577-021-00522-1
发表时间：
2021-04
期刊：
Nature reviews. Immunology
影响因子：
--
作者：
Carvalho T;Krammer F;Iwasaki A
通讯作者：
Iwasaki A
影响因子：
3.1
作者：
Sharma, Sai Kiran;Lyashchenko, Serge K.;Lewis, Jason S.
通讯作者：
Lewis, Jason S.
影响因子：
14.2
作者：
To, K. K. W.;Yip, C. C. Y.;Yuen, K-Y
通讯作者：
Yuen, K-Y
影响因子：
2.4
作者：
Xu, Rongbin;Mei, Yang;Liu, Jianping
通讯作者：
Liu, Jianping