Evaluation of SARS-CoV-2 isolation in cell culture from nasal/nasopharyngeal swabs or saliva specimens of patients with COVID-19

Yazawa et al., Research Square, doi:10.21203/rs.3.rs-2676422/v1 (Preprint) (In Vitro)

Yazawa et al., Evaluation of SARS-CoV-2 isolation in cell culture from nasal/nasopharyngeal swabs or saliva specimens of.., Research Square, doi:10.21203/rs.3.rs-2676422/v1 (Preprint) (In Vitro)

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Analysis of nasal/nasopharyngeal swabs and saliva specimens, showing significantly lower isolation efficiency of SARS-CoV-2 in saliva specimens. In Vitro study showing that lactoferrin and amylase, components of saliva, were found to inhibit SARS-CoV-2 pseudotyped virus infection.

7 In Vitro studies support the efficacy of lactoferrin [Andreu, Cutone, Mirabelli, Ostrov, Piacentini, Salaris, Yazawa].

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1. Andreu et al., Viruses, doi:10.3390/v15040972, Liposomal Lactoferrin Exerts Antiviral Activity against HCoV-229E and SARS-CoV-2 Pseudoviruses In Vitro, https://www.mdpi.com/1999-4915/15/4/972.

2. Cutone et al., Research Square, doi:10.21203/rs.3.rs-1605740/v1, Lactoferrin binding to Sars-CoV-2 Spike glycoprotein protects host from infection, inflammation and iron dysregulation., https://www.researchsquare.com/article/rs-1605740/v1.

3. Mirabelli et al., Proceedings of the National Academy of Sciences, doi:10.1073/pnas.2105815118, Morphological cell profiling of SARS-CoV-2 infection identifies drug repurposing candidates for COVID-19, https://www.pnas.org/content/118/36/e2105815118.

4. Ostrov et al., Pathogens, doi:10.3390/pathogens10111514, Highly Specific Sigma Receptor Ligands Exhibit Anti-Viral Properties in SARS-CoV-2 Infected Cells, https://www.mdpi.com/2076-0817/10/11/1514/htm.

5. Piacentini et al., International Journal of Molecular Sciences, doi:10.3390/ijms23105436, Lactoferrin Inhibition of the Complex Formation between ACE2 Receptor and SARS CoV-2 Recognition Binding Domain, https://www.mdpi.com/1422-0067/23/10/5436.

6. Salaris et al., Nutrients, doi:10.3390/nu13020328, Protective Effects of Lactoferrin against SARS-CoV-2 Infection In Vitro, https://www.mdpi.com/2072-6643/13/2/328.

7. Yazawa et al., Research Square, doi:10.21203/rs.3.rs-2676422/v1, Evaluation of SARS-CoV-2 isolation in cell culture from nasal/nasopharyngeal swabs or saliva specimens of patients with COVID-19, https://www.researchsquare.com/article/rs-2676422/v1.

Yazawa et al., 31 Mar 2023, preprint, 8 authors.

Contact: toyamaeiken3@juno.ocn.ne.jp.

In Vitro studies are an important part of preclinical research, however results may be very different in vivo.

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Evaluation of SARS-CoV-2 isolation in cell culture from nasal/nasopharyngeal swabs or saliva specimens of patients with COVID-19

Shunsuke Yazawa, Emiko Yamazaki, Yumiko Saga, Masae Itamochi, Noriko Inasaki, Takahisa Shimada, Kazunori Oishi, Hideki Tani

doi:10.21203/rs.3.rs-2676422/v1

It has been revealed that SARS-CoV-2 can be e ciently isolated from clinical specimens such as nasal/nasopharyngeal swabs or saliva in cultured cells. In this study, we examined the e ciency of viral isolation including SARS-CoV-2 mutant strains between nasal/nasopharyngeal swab or saliva specimens. Furthermore, we also examined the comparison of viral isolation rates by sample species using simulated specimens for COVID-19. As a result, it was found that the isolation e ciency of SARS-CoV-2 in the saliva specimens was signi cantly lower than that in the nasal/nasopharyngeal swab specimens. In order to determine which component of saliva is responsible for the lower isolation rate of saliva specimens, we tested the abilities of lactoferrin, amylase, cathelicidin, and mucin, which are considered to be abundant in saliva, to inhibit the infection of SARS-CoV-2 pseudotyped viruses (SARS-CoV-2pv). Lactoferrin and amylase were found to inhibit SARS-CoV-2pv infection. In conclusion, even if the same number of viral genome copies was detected by the real-time RT-PCR test, infection of SARS-CoV-2 present in saliva is thought to be inhibited by inhibitory factors such as lactoferrin and amylase, compared to nasal/nasopharyngeal swab specimens.

Declarations Ethical approval This study was performed in accordance with the Helsinki Declaration and was approved by the ethical review board of the Toyama Institute of Health (approval No.: R2-1). The need of informed consent was also waived by the ethical review board of the Toyama Institute of Health (approval No.: R2-1). Con ict of interest We have no potential con icts of interest in relation to this work.

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