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Review 1: "Development of a Simple and Highly Sensitive Virion Concentration Method to Detect SARS-CoV-2 in Saliva"

The reviewers found the hypothesis to be sound and the data reliable. However, they raised concerns about the practicality of using 12ml saliva samples, which is a large volume and not attainable for many patients.

Published onJun 06, 2024
Review 1: "Development of a Simple and Highly Sensitive Virion Concentration Method to Detect SARS-CoV-2 in Saliva"
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key-enterThis Pub is a Review of
Development of a simple and highly sensitive virion concentration method to detect SARS-CoV-2 in saliva
Development of a simple and highly sensitive virion concentration method to detect SARS-CoV-2 in saliva
Description

Abstract Background Controlling novel coronavirus pandemic infection (COVID-19) is a global challenge, and highly sensitive testing is essential for effective control. The saliva is a promising sample for high-sensitivity testing because it is easier to collect than nasopharyngeal swab samples and allows large-volume testing.Results We developed a simple SARS-CoV-2 concentration method from saliva samples that can be completed in less than 60 min. We performed a spike test using 12 ml of saliva samples obtained from healthy volunteer people, and the developed method performance was evaluated by comparison using a combination of automatic nucleic acid extraction followed by RT-qPCR detection. In saliva spike tests using a 10-fold dilution series of SARS-CoV-2, the developed method was consistently 100-fold more sensitive than the conventional method.Conclusions The developed method can improve the sensitivity of the SARS-CoV-2 test using saliva and speed up and save labor in screening tests by pooling many samples. Furthermore, the developed method has the potential to contribute to the highly sensitive detection of various human and animal viral pathogens from the saliva and various clinical samples.Highlight A method has been developed to detect SARS-CoV-2 from human saliva with 100 times higher sensitivity than conventional methods.The developed method combines simple pretreatment within 60 min with conventional nucleic acid extraction and RT-qPCR.This method can be applied for more sensitive virus testing from individual saliva.This method can potentially be applied to screening more than 100 saliva samples while maintaining the equivalent detection power of conventional methods.The method can be adapted to improve the sensitivity of detecting various pathogens from human and animal saliva.

RR:C19 Evidence Scale rating by reviewer:

  • Potentially informative. The main claims made are not strongly justified by the methods and data, but may yield some insight. The results and conclusions of the study may resemble those from the hypothetical ideal study, but there is substantial room for doubt. Decision-makers should consider this evidence only with a thorough understanding of its weaknesses, alongside other evidence and theory. Decision-makers should not consider this actionable, unless the weaknesses are clearly understood and there is other theory and evidence to further support it.

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Review: The paper is well written and appears to be reproducable in other labs. It is mainly a technique paper. By starting with a larger sample (rather large by the way and practicality of using 12ml of saliva sample can be questioned), and adding additional preconcentration steps, the authors claim that the LOD can be improved by 100 times. The hypothesis makes sense - if you start with larger sample and indeed preconcencrate more, clearly enough viruses can be retained in the final samples. The data appears to be reliable. As a technqiue paper this a small, solid, and perhaps useful addition to the literature. As noted above the use of a 12ml of saliva is not trivial and cannot be provided by all patients and specially children or the elderly. So this is clearly only viable as a technique for control measurements where large amount of saliva is available. 

References that could be added:

  1. D. R. E. Ranoa, et al., "Mitigation of SARS-CoV-2 Transmission at a Large Public University", Nat. Comm. 13, 3207 (2022); doi:10.1038/s41467-022-30833-3.

  2. A. Ganguli, A. Mostafa, J. Berger, J. Lim, E. Araud, J. Baek, S. A. Stewart de Ramirez, A. Baltaji, K. Roth, M. Aamir, S. Aedma, M. Mady, P. Mahajan, S. Sathe, M. Johnson, K. White, J. Kumar, E. Valera, R. Bashir, "Reverse Transcription Loop-Mediated Isothermal Amplification Assay for Ultrasensitive Detection of SARS-CoV-2 in Saliva and Viral Trnsport Medium Clinical Samples", Anal. Chem., 93, 7797, (2021); doi:10.1021/acs.analchem.0c05170.

  3. A. Ganguli, A. Mostafa, J. Berger, M. Y. Aydin, F. Sun, S. A. S. de Ramirez, E. Valera, B. T. Cunningham, W. P. King, and R. Bashir, "Rapid isothermal amplification and portable detection system for SARS-CoV-2", PNAS, 117, 22727 (2020); doi:10.1073/pnas.2014739117.

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