Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients

Igarashi, Emiko; Tani, Hideki; Tamura, Kosuke; Itamochi, Masae; Shimada, Takahisa; Saga, Yumiko; Inasaki, Noriko; Hasegawa, Sumiyo; Yazawa, Shunsuke; Sasajima, Hitoshi; Kaya, Hiroyasu; Nomura, Satoshi; Itoh, Hiroyuki; Takeda, Shin‐ichi; Yamamoto, Yoshihiro; Yamashiro, Seiji; Ichikawa, Tomomi; Horie, Yukio; Hirota, Kojiro; Hirano, Naoki; Kawashiri, Chikako; Oishi, Kazunori

doi:10.1016/j.jiac.2021.10.028

“…Nasal/nasopharyngeal swabs or saliva specimens were stored at − 80 °C before being processed in cell culture. VeroE6/TMPRSS2 cells were inoculated with nasal/nasopharyngeal swabs or saliva specimens as described in an earlier report by Igarashi et al 3 . Briefly, 20 μL of each nasal/nasopharyngeal swabs or saliva cleaning solution, which exhibited rRT-PCR positivity for SARS-CoV-2, was added to VeroE6/TMPRSS2 cells seeded the preceding day on a 24-well plate and cultured at 37 °C for 5 days, and the cytopathic effect (CPE) was confirmed by visual observation under a microscope.…”

Section: Methodsmentioning

confidence: 99%

“…We previously evaluated the viral isolation rate using cultured cells from clinical specimens positive for SARS-CoV-2, and analyzed the correlation with the Ct value of the rRT-PCR test 3 . However, although virus was efficiently isolated from the nasal/nasopharyngeal swab specimens, many copies could not be isolated from the saliva specimens.…”

Section: Introductionmentioning

confidence: 99%

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

Yazawa

¹

,

Yamazaki

²

,

Saga

³

et al. 2023

Sci Rep

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It has been revealed that SARS-CoV-2 can be efficiently isolated from clinical specimens such as nasal/nasopharyngeal swabs or saliva in cultured cells. In this study, we examined the efficiency 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 efficiency of SARS-CoV-2 in the saliva specimens was significantly 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.

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“…Nasal/nasopharyngeal swabs or saliva specimens were stored at -80˚C before being processed in cell culture. VeroE6/TMPRSS2 cells were inoculated with nasal/nasopharyngeal swabs or saliva specimens as described in an earlier report by Igarashi et al 3 . Brie y, 20 µL of each nasal/nasopharyngeal swabs or saliva cleaning solution, which exhibited rRT-PCR positivity for SARS-CoV-2, was added to VeroE6/TMPRSS2 cells seeded the preceding day on a 24-well plate and cultured at 37°C for ve days, and the cytopathic effect (CPE) was con rmed by visual observation under a microscope.…”

Section: Viral Isolation Test From Clinical Specimensmentioning

confidence: 99%

“…We previously evaluated the viral isolation rate using cultured cells from clinical specimens positive for SARS-CoV-2, and analyzed the correlation with the Ct value of the rRT-PCR test 3 . However, a though virus was e ciently isolated from the nasal/nasopharyngeal swab specimens, many copies could not be isolated from the saliva specimens.…”

Section: Introductionmentioning

confidence: 99%

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

Yazawa

¹

,

Yamazaki

²

,

Saga

³

et al. 2023

Preprint

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0

View full text Add to dashboard Cite

It has been revealed that SARS-CoV-2 can be efficiently isolated from clinical specimens such as nasal/nasopharyngeal swabs or saliva in cultured cells. In this study, we examined the efficiency 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 efficiency of SARS-CoV-2 in the saliva specimens was significantly 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.

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“…Different reports suggest lower Ct values associated with higher viral load, viral recovery and more severe illness; therefore, this may aid in the pretransplant evaluation. 5 , 6 The values that have shown to not be associated with cell culture viral recovery or range from >24 (especially after 8 days from symptom onset) to 34. 5 , 7 , 8 We present two patients who underwent successful deceased donor liver transplantation (DDLT) with a positive SARS‐CoV‐2 nasopharyngeal PCR.…”

mentioning

confidence: 99%

Successful deceased donor liver transplantation in candidates with high MELD and ongoing positive SARS‐CoV‐2 PCR

Vissichelli

¹

,

Morales

²

,

Bryson

³

et al. 2022

Transplant Infectious Dis

1

0

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Viral isolation analysis of SARS-CoV-2 from clinical specimens of COVID-19 patients

Cited by 8 publications

References 8 publications

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

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

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

Successful deceased donor liver transplantation in candidates with high MELD and ongoing positive SARS‐CoV‐2 PCR

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