Review of Paper Methods: Isolating the COVID-19 "Virus"

In this video I go over further into the main paper on COVID-19 "virus" isolation, and this time review the paper's methods in isolating the "novel virus". The stated methods involved first collecting the lung fluid samples in a plastic cup, and then chemical preservatives (PBS) or fetal bovine serum (FBS) were added. The samples were then centrifuged and the resulting supernatant was assumed to somehow contain the "virus" and used to "infect" human airway cell cultures obtained from other lung cancer patients. Then the cultures were "incubated" or grown for 2 hours at 37°C, and then diluted to a new cell culture. This procedure was done 3 times total and each time the cell cultures were monitored using optical light microscopy for "cytopathic effects" (or cell breakdown / death). Cultures showing cytopathic effects were then prepared for electron microscopy. No information on the procedure for controls were presented. And no explanation as to why the centrifuged supernatant of the patient lung fluid samples were assumed to contain the "virus".

The paper is listed below:

• A Novel Coronavirus from Patients with Pneumonia in China, 2019
• Authors: Na Zhu et al
• Published on January 24, 2020 in The New England Journal of Medicine (NEJM)
• https://www.nejm.org/doi/full/10.1056/NEJMoa2001017

The timestamps of key parts of the video are listed below:

• Paper Methods: Isolating the "Virus": 0:00
• Viral Transport Medium (VTM), Acids and Bases : 1:14
• Fetal Bovine Serum (FBS): 3:08
• Samples Centrifuged, Supernatant Use to Infect: 3:57
• Centrifugation, Supernatant, Cancer: 4:31
• Cells Grown on Plastic Surface: 7:01
• Plastic substrate, Expanding Cells, Passage-1 Cells, Plating: 7:29
• Cells Cultures Grown in Air-Liquid Interface: 9:42
• Air-Liquid Interface (ALI), Cellular Differentiation: 10:04
• Stem Cells, Respiratory Epithelium, Cilia: 12:57
• Apical Cells Washed with PBS: 14:14
• Apical, Microlitre: 15:08
• MES Note: Where is the "Virus"?: 16:13
• 2-hour Incubation, Unbound "Virus" Washed Away: 16:40
• Incubator, 37° C, Serial Passage: Cultural Dilution: 18:16
• Cells Monitored for Cytopathic Effects: 19:12
• Light Microscopy, Cytopathic Effect: 19:54
• Summary of Isolating the "Virus": 20:34

This video is taken from my earlier video listed below:

• 🔥#MESScience 4: Review of COVID-19 "Virus" Isolation Paper: https://youtu.be/cvDO85Rw4d8
• Video notes: https://peakd.com/hive-128780/@mes/messcience-4-review-of-covid-19-virus-isolation-paper
• Playlist: https://www.youtube.com/playlist?list=PLai3U8-WIK0E4aQ_cq4ZDD2WGiDU5vVgx

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