| Eunbyeol Wang | 2 Articles |
<sec><title>Objectives</title>
<p>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.</p></sec>
<sec><title>Methods</title>
<p>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.</p></sec>
<sec><title>Results</title>
<p>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.</p></sec>
<sec><title>Conclusion</title>
<p>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.</p></sec>
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Objectives
The purpose of this study was to verify the feasibility of using the glyceraldehyde-3-phosphate dehydrogenase (GAP) promotor based <i>Pichia pastoris</i> 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 <i>P. pastoris</i>, 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 <i>P. pastoris</i> expression system. The results of Western blotting showed that the recombinant proteins were secreted into the culture supernatant from the <i>P. pastoris</i> and glycosylated. Conclusion This study suggests that recombinant TBEV VLPs from <i>P. pastoris</i> offer a promising approach to the production of VLPs for use as vaccines and diagnostic antigens. Citations
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