I am giving you a link to my doctoral dissertation. I isolated a previously unknown Spumavirinae and you can look up in the experimental methods the how to do serial passage of live virus in tissue cultures.

https://archive.org/stream/isolationcharact00laha#page/n41/mode/2up

As per WHO recommendation BSL3 level lab is required for SARS-CoV-19 lab culture. If you have never done viral culture please do NOT do it without proper training. It is dangerous.

However, for animal infectivity you have to talk to experienced virologist as the procedure is too technical that you require proper training.

See the WHO guide

https://www.who.int/docs/default-source/coronaviruse/laboratory-biosafety-novel-coronavirus-version-1-1.pdf?sfvrsn=912a9847_2

If you will do cytometry analysis please refer to "Handling aand Processing of Blood Specimens from patients with COVID 19for Safe Studies on Cell Phenotype and Cytokine Storm" published by A. Cossarizza et al in Cytometry A.

Best wishes,

Not sure why Jiang Kunpeng says you need BSL4. You need CL3 for SARS-CoV-2, if it's an area you want to work in I'd approach someone with an already established CL3 facility, you need a lot of paperwork and approval in most countries before it can be used at CL3. We're converting a TB CL3 lab to work with SARS-CoV-2 and it took us a few weeks to get permission. However, when we first setup the TB lab it took many months to get all the approvals in place

Sion Robert Phillips,

where is your lab located. I would very much like to collaborate.

1) This morning I again saw a doctor on CNN question the safety of chloroquine. But a billion people living in lands where malaria is endemic have taken it. Chloroquine is a life saver. It has prevented millions of death. It’s been taken by tens of millions on a weekly basis for prevention, I for one, or daily if afflicted with malaria. So what is the safety issue?

2) Testing its efficacy on inhibiting viral growth in ciliated tracheal and naso/pharyngeal cell cultures should have been completed long ago.

3) Large representative samples of the population should have already been tested with simple serum antibody precipitation test for IgG or IgM and similar swab tests for IgA(the secretory antibody) using the isolated Spike protein from the virus as antigen. This is the only way to really estimate the real % of the population infected and the real morbidity/mortality rate of COViD19. But instead an expensive RT/PRC test was being done and only for people with symptoms which can never give us these answers.

4) A vaccine using the receptor site protein of the virus in an appropriate adjuvant should have already been tested on animals and human trials in progress.

I say these things with confidence because I did those procedures since 1978 with a new virus I isolated in the viral family Retroviridae with the rudimentary technology available then. Isolation of the virus, experimental infection of laboratory animals, production of antisera, conjugation of antibodies with FITC to illuminate the virus, and immunodiffusion tests on large numbers of sera to see how endemic it was. So i am totally perplexed why such steps have not yet been accomplished 43 years later with this virus.

certainly BSL-3 facility

https://www.cdc.gov/sars/guidance/f-lab/app5.html

Hello every body.

The firts thing that we do is know that it is necesarry a BSL-3 level for working with coronavirus SARS-CoV 2 culture, because the production of aerosols.

The second thing is very very important the workers qualification in BSL-3 work.

I have worked a lot of time with Guanarito virus, a virus classified by OMS Risk 4, and I work with culture viral.But it is necessary have experience, training, competency in viral culture.Avoid the LAIS

Yes , on verocell line to check the adhesion of virus on cell.

For me, It is difficult because COVID 19 is a new virus

The first Corona viruses were isolated from embryonic ciliate tracheal cell and nasopharyngeal/pharingea cell cultures. Those would be the best cultures to grow the Coronas and test what inhibits such growth.

Actually our lab in Siena (Italy) is working on it in our BSL3 facility.

Thanks to everyone responded! However, you did not quite understand me correctly. We have experience in operating with rabies virus and test the antiviral activity of drugs on it , there are also some clinical trials on other pathogens (malaria and HIV). However, we do not going to equip our laboratory to work with Sars-cov-2 (we have BSL 2). We would like to provide our substance to laboratories that are already working with this pathogen for testing against new coronavirus.