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Ye-Ji Lee 3 Articles
Detection and Isolation of SARS-CoV-2 in Serum, Urine, and Stool Specimens of COVID-19 Patients from the Republic of Korea
Jeong-Min Kim, Heui Man Kim, Eun Jung Lee, Hye Jun Jo, Youngsil Yoon, Nam-Joo Lee, Junseock Son, Ye-Ji Lee, Mi Seon Kim, Yong-Pyo Lee, Su-Jin Chae, Kye Ryeong Park, Seung-Rye Cho, Sehee Park, Su Jin Kim, Eunbyeol Wang, SangHee Woo, Aram Lim, Su-Jin Park, JunHyeong Jang, Yoon-Seok Chung, Bum Sik Chin, Jin-Soo Lee, Duko Lim, Myung-Guk Han, Cheon Kwon Yoo
Osong Public Health Res Perspect. 2020;11(3):112-117.   Published online May 8, 2020
DOI: https://doi.org/10.24171/j.phrp.2020.11.3.02
  • 9,585 View
  • 502 Download
  • 74 Citations
AbstractAbstract PDF
Objectives

Coronavirus Disease-19 (COVID-19) is a respiratory infection characterized by the main symptoms of pneumonia and fever. It is caused by the novel coronavirus severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2), which is known to spread via respiratory droplets. We aimed to determine the rate and likelihood of SARS-CoV-2 transmission from COVID-19 patients through non-respiratory routes.

Methods

Serum, urine, and stool samples were collected from 74 hospitalized patients diagnosed with COVID-19 based on the detection of SARS-CoV-2 in respiratory samples. The SARS-CoV-2 RNA genome was extracted from each specimen and real-time reverse transcription polymerase chain reaction performed. CaCo-2 cells were inoculated with the specimens containing the SARS-COV-2 genome, and subcultured for virus isolation. After culturing, viral replication in the cell supernatant was assessed.

Results

Of the samples collected from 74 COVID-19 patients, SARS-CoV-2 was detected in 15 serum, urine, or stool samples. The virus detection rate in the serum, urine, and stool samples were 2.8% (9/323), 0.8% (2/247), and 10.1% (13/129), and the mean viral load was 1,210 ± 1,861, 79 ± 30, and 3,176 ± 7,208 copy/µL, respectively. However, the SARS-CoV-2 was not isolated by the culture method from the samples that tested positive for the SARS-CoV-2 gene.

Conclusion

While the virus remained detectable in the respiratory samples of COVID-19 patients for several days after hospitalization, its detection in the serum, urine, and stool samples was intermittent. Since the virus could not be isolated from the SARS-COV-2-positive samples, the risk of viral transmission via stool and urine is expected to be low.

One-Step Reverse Transcription-Polymerase Chain Reaction for Ebola and Marburg Viruses
Sun-Whan Park, Ye-Ji Lee, Won-Ja Lee, Youngmee Jee, WooYoung Choi
Osong Public Health Res Perspect. 2016;7(3):205-209.   Published online June 30, 2016
DOI: https://doi.org/10.1016/j.phrp.2016.04.004
  • 1,405 View
  • 23 Download
  • 4 Citations
AbstractAbstract PDF
Objectives
Ebola and Marburg viruses (EBOVs and MARVs, respectively) are causative agents of severe hemorrhagic fever with high mortality rates in humans and nonhuman primates. In 2014, there was a major Ebola outbreak in various countries in West Africa, including Guinea, Liberia, Republic of Sierra Leone, and Nigeria. EBOV and MARV are clinically difficult to diagnose and distinguish from other African epidemic diseases. Therefore, in this study, we aimed to develop a method for rapid identification of the virus to prevent the spread of infection.
Methods
We established a conventional one-step reverse transcription-polymerase chain reaction (RT-PCR) assay for these pathogens based on the Superscript Reverse Transcriptase-Platinum Taq polymerase enzyme mixture. All assays were thoroughly optimized using in vitro-transcribed RNA.
Results
We designed seven primer sets of nucleocapsid protein (NP) genes based on sequences from seven filoviruses, including five EBOVs and two MARVs. To evaluate the sensitivity of the RT-PCR assay for each filovirus, 10-fold serial dilutions of synthetic viral RNA transcripts of EBOV or MARV NP genes were used to assess detection limits of viral RNA copies. The potential for these primers to cross react with other filoviruses was also examined. The results showed that the primers were specific for individual genotype detection in the examined filoviruses.
Conclusion
The assay established in this study may facilitate rapid, reliable laboratory diagnosis in suspected cases of Ebola and Marburg hemorrhagic fevers.
Cloning and Expression of Recombinant Tick-Borne Encephalitis Virus-like Particles in Pichia pastoris
Seok-Min Yun, Young Eui Jeong, Eunbyeol Wang, Ye-Ji Lee, Myung Guk Han, Chan Park, Won-Ja Lee, WooYoung Choi
Osong Public Health Res Perspect. 2014;5(5):274-278.   Published online October 31, 2014
DOI: https://doi.org/10.1016/j.phrp.2014.08.005
  • 1,505 View
  • 16 Download
  • 7 Citations
AbstractAbstract PDF
Objectives
The purpose of this study was to verify the feasibility of using the glyceraldehyde-3-phosphate dehydrogenase (GAP) promotor based Pichia pastoris expression system to produce tick-borne encephalitis virus (TBEV) virus-like particles (VLPs).
Methods
The complementary DNA encoding the TBEV prM signal peptide, prM, and E proteins of TBEV Korean strain (KrM 93) was cloned into the plasmid vector pGAPZɑA, then integrated into the genome of P. pastoris, under the control of the GAP promoter. Expression of TBEV VLPs was determined by Western blotting using monoclonal antibody against TBEV envelope (E) protein.
Results
Recombinant TBEV VLPs consisting of prM and E protein were successfully expressed using the GAP promoter-based P. pastoris expression system. The results of Western blotting showed that the recombinant proteins were secreted into the culture supernatant from the P. pastoris and glycosylated.
Conclusion
This study suggests that recombinant TBEV VLPs from P. pastoris offer a promising approach to the production of VLPs for use as vaccines and diagnostic antigens.

PHRP : Osong Public Health and Research Perspectives