Recently, employees of Rowpar Pharmaceuticals Inc. wrote a commentary entitled, “Overinterpretation of the antiviral results for human coronavirus 229E (HCoV‐229E) relative to severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2).” In their commentary, they discussed our recent publication entitled, “Lower the transmission and spread of human coronavirus.” In their commentary, there is a limited discussion of accepted virucidal analysis, confusing virucidal assays with assays to determine mechanisms of infection. Also ignored is the corroboration of our studies by others using SARS‐CoV‐2, including a clinical trial. We will now address their comments, including providing the corroborative published studies.

First, our published studies measure the efficacy of nasal and oral rinses to inactivate HCoV, representing the beginning stage of identifying a process whereby transmission and spread can be lowered. We also conclude the studies by saying, “While clinical trials will be necessary to confirm the virucidal potential of these products and assess their ability to limit transmission of HCoV within the general population, in the current manuscript we have demonstrated here that several commonly available healthcare products have significant virucidal properties with respect to HCoV.”

Second, it is quite common and acceptable in the disinfection/virucidal field to use surrogate viruses, particularly members of the same virus family. One clear example of this is the list provided by the Environmental Protection Agency “Disinfectants for Use Against SARS‐CoV‐2,”¹ which lists hundreds of acceptable virucidal agents based on their efficacy using surrogate HCoV, other nonrelated enveloped and nonenveloped virus, using numerous types of cell lines, and contact times. However, it is always desired that eventually the studies using viral surrogates can be corroborated using the actual infectious agent of concern. At the same time as we were doing our studies, three other research teams were performing similar studies using SARS‐CoV‐2 and reported similar results as we had found in our study. The first two were available online when we published and are included in the discussion of our manuscript. Both demonstrated that povidone–iodine solutions are highly effective at inactivating SARS‐CoV‐2^{2, 3} and one demonstrated that hydrogen peroxide solutions were only minimally effective.² These results are the same results reported in our manuscript. The third research team demonstrated that commercially available oral rinses, including listerine, efficiently inactivated SARS‐CoV‐2.⁴ Again, these results are the same results reported in our manuscript. Therefore, our conclusions do not contradict extant literature as suggested by Rowpar Pharmaceuticals. In addition, numerous clinical trials have been started to test the efficacy of nasal and oral rinses to lower the viral load of coronavirus disease 2019 patients with 15 separate trials listed on ClinicalTrials.gov. One small trial has already been completed and found that listerine lowered viral load by 80%.⁵

Third, there is confusion in Rowpar Pharmaceuticals' comments concerning assaying for a disinfectant of the virucidal compound for its ability to inactivate infectious virus as opposed to studying the mechanisms of infectivity. For example, Rowpar Pharmaceuticals discusses the specificity of the SARS‐CoV‐2 spike protein to binding the cell angiotensin‐converting enzyme 2 receptor and the structural differences between the SARS‐CoV‐2 spike protein and the SARS‐CoV spike protein. While definitely important for the mechanism of infectivity and vaccine design, it has little if anything to do with the ability of a virucidal agent to disrupt the viral envelope.

Fourth, Rowpar Pharmaceuticals states that “even among different strains of SARS‐CoV‐2” there are differences in the efficacy of inactivation. In one study, the reference points out log₁₀ decreases in infectivity for three strains as ≥3.11, >2.78, and ≥2.61 for strains 1–3, respectively.⁴ We completely agree that tests and comparison should be done with variants of SARS‐CoV‐2. However, these values are not statistically different and the authors state, “We found that different SARS‐CoV‐2 strains can be efficiently inactivated with commercially available oral rinses under biologically relevant conditions mimicking respiratory secretions” and “In agreement with our observation, different studies using Listerine (product F) observed antiviral activities specifically against enveloped viruses, implying an impact on the viral lipid envelope.”⁴ Of the two other examples mentioned by Rowpar Pharmaceuticals, the first compares SARS‐CoV‐2 against Influenza A virus and even a nonenveloped virus human rotavirus. The second compares numerous viruses enveloped and nonenveloped, none of which is even a coronavirus. Rowpar Pharmaceuticals also point out in Table 1 of their commentary that their own product, CloSYS Ultra Sensitive Rinse, which while effective at inactivating influenza A virus is ineffective at inactivating SARS‐CoV‐2.

In summary, in our experimental design, we followed standard accepted practices in assaying virucidal activity. Results were not overinterpreted as we clearly stated the possible limitations of our study in the final paragraph of our Discussion. Our findings did not contradict the extant literature or studies that have been published afterward and have been corroborated using SARS‐CoV‐2. Our work along with the excellent work of others has provided a strong foundation for the continued study of nasal and oral rinses in lowering the SARS‐CoV‐2 transmission, including clinical trials.

最近，Rowpar Pharmaceuticals Inc.的员工写了一篇题为“对人冠状病毒229E（HCoV-229E）相对于严重急性呼吸系统综合症冠状病毒2（SARS-CoV-2）的抗病毒结果的过度解释”的评论。他们在评论中讨论了我们最近的出版物，题为“降低人类冠状病毒的传播和传播”。在他们的评论中，对接受的杀病毒分析进行了有限的讨论，将杀病毒分析与确定感染机理的分析相混淆。也被其他人使用SARS-CoV-2的研究证实，包括一项临床试验，也被忽略了。现在，我们将解决他们的评论，包括提供证实性的已发表研究。

首先，我们发表的研究测量了鼻腔冲洗和口服漱口水使HCoV失活的功效，这代表了确定可降低传播和传播过程的开始阶段。我们还以以下结论结束了研究：“虽然需要进行临床试验来确认这些产品的杀病毒潜力，并评估其限制普通人群中HCoV传播的能力，但在目前的手稿中，我们在此处证明了几种常用的医疗保健方法产品具有相对于HCoV的显着的杀病毒性能。”

第二，使用替代病毒，尤其是同一病毒家族的成员，在消毒/杀病毒领域是很普遍且可以接受的。一个明显的例子是环境保护局提供的“用于对抗SARS-CoV-2的消毒剂”清单¹根据使用替代HCoV，其他不相关的包膜和非包膜病毒，使用多种类型的细胞系和接触时间的功效列出了数百种可接受的杀病毒剂。但是，始终希望最终可以使用实际关注的传染因子来证实使用病毒替代物的研究。在进行研究的同时，其他三个研究小组也使用SARS-CoV-2进行了相似的研究，并报告了与研究中相似的结果。前两个版本在我们发布时可以在线获得，并包含在我们手稿的讨论中。既表明，聚维酮碘溶液是在灭活SARS-CoV的-2高效^2,3有人证明过氧化氢溶液的作用很小。²这些结果与我们手稿中报道的结果相同。第三个研究小组证明，市售的口腔冲洗剂（包括李斯汀）可以有效地灭活SARS-CoV-2。⁴同样，这些结果与我们手稿中报道的结果相同。因此，我们的结论与Rowpar Pharmaceuticals建议的现有文献并不矛盾。此外，许多临床试验已开始进行临床试验，以临床和临床试验中列出的15项单独试验来测试鼻腔冲洗和口服漱口水降低2019年冠状病毒病患者病毒载量的功效。一项小型试验已经完成，发现李斯汀可将病毒载量降低80％。⁵

第三，在Rowpar Pharmaceuticals的评论中，关于研究杀病毒化合物消毒剂的灭活传染性病毒的能力，而不是研究传染性机理，存在混淆。例如，Rowpar Pharmaceuticals讨论了SARS-CoV-2刺突蛋白与细胞血管紧张素转化酶2受体结合的特异性，以及SARS-CoV-2刺突蛋白与SARS-CoV刺突蛋白之间的结构差异。虽然对于感染性和疫苗设计的机制绝对重要，但与杀病毒剂破坏病毒包膜的能力几乎没有关系。

第四，Rowpar Pharmaceuticals指出“即使在SARS-CoV-2的不同菌株之间”，灭活效果也存在差异。在一项研究中，参考文献指出三种菌株的传染性log ₁₀降低，分别为菌株1-3≥3.11，> 2.78和≥2.61。⁴我们完全同意应使用SARS-CoV-2的变体进行测试和比较。但是，这些值在统计学上没有差异，作者指出：“我们发现，可以在与呼吸道分泌相似的生物学相关条件下，使用市售的口腔冲洗液有效地灭活不同的SARS-CoV-2菌株”和“与我们的观察一致，不同使用Listerine（产品F）的研究观察到了专门针对包膜病毒的抗病毒活性，这暗示了对病毒脂质包膜的影响。” ⁴在Rowpar Pharmaceuticals提到的另外两个例子中，第一个将SARS-CoV-2与A型流感病毒甚至非包膜病毒人轮状病毒进行了比较。第二种比较了许多包膜和非包膜的病毒，甚至没有一种是冠状病毒。Rowpar Pharmaceuticals在其评论的表1中还指出，他们自己的产品CloSYS Ultra Sensitive Rinse，虽然可以有效灭活A型流感病毒，但不能有效灭活SARS-CoV-2。

总之，在我们的实验设计中，我们遵循标准的公认实践来测定杀病毒活性。由于我们在讨论的最后一段中明确指出了研究的可能局限性，因此并未对结果进行过分解释。我们的发现与随后发表的并使用SARS-CoV-2证实的现有文献或研究没有矛盾。我们的工作以及其他人的出色工作为继续研究鼻腔和口腔冲洗液以降低SARS-CoV-2传播（包括临床试验）奠定了坚实的基础。