[minit 18:27] Patents by Centers for Disease Control and Prevention/CDC that cover the gene sequence of the coronavirus and the means of detecting it using RT-PCR
[minit 27:55] United States Department of Defense/DARPA actively develop coronavirus as biological weapon - SARS CoV-2
05.06.2008 Application
24.05.2015 Patent US9193780 Amino acid sequences directed against envelope proteins of a virus and polypeptides comprising the same for the treatment of viral diseases [PDF]
13.10.2015 : US Patent US20200279585A1 : "A method is provided for acquiring and transmitting biometric data (e.g., vital signs) of a user, where the data is analyzed to determine whether the user is suffering from a viral infection, such as COVID-19," System and Method for Testing for COVID-19 filed by Richard Rothchild. Refer to 11.02.2020.
The RNA and GENE strands / of the SARS CoV-2 SPIKE PROTEIN - 73 Patents Issued
09.01.2020
Chinese authorities have determined that the outbreak is caused by a novel coronavirus.
ChinaNews.com : “The discovery of nucleic acid, genomic and antibody evidence of the pathogen from patients can be accomplished in a short period of time. Scientific studies such as the isolation and pathogenicity identification of the pathogen can take several weeks.”
16-24.02.2020 WHO Report of the WHO-China Joint Mission on Coronavirus Disease 2019 (COVID-19)
"The virus
On 30 December 2019, three bronchoalveolar lavage samples were collected from a patient with pneumonia of unknown etiology – a surveillance definition established following the SARS outbreak of 2002-2003 – in Wuhan Jinyintan Hospital. Real-time PCR (RT-PCR) assays on these samples were positive for pan-Betacoronavirus. Using Illumina and nanopore sequencing, the whole genome sequences of the virus were acquired. Bioinformatic analyses indicated that the virus had features typical of the coronavirus family and belonged to the Betacoronavirus 2B lineage. Alignment of the full-length genome sequence of the COVID-19 virus and other available genomes of Betacoronavirus showed the closest relationship was with the bat SARS-like coronavirus strain BatCov RaTG13, identity 96%.
Virus isolation was conducted with various cell lines, such as human airway epithelial cells, Vero E6, and Huh-7. Cytopathic effects (CPE) were observed 96 hours after inoculation. Typical crown-like particles were observed under transmission electron microscope (TEM) with negative staining. The cellular infectivity of the isolated viruses could be completely neutralized by the sera collected from convalescent patients. Transgenic human ACE2 mice and Rhesus monkey intranasally challenged by this virus isolate induced multifocal pneumonia with interstitial hyperplasia. The COVID-19 virus was subsequently detected and isolated in the lung and intestinal tissues of the challenged animals.
Whole genome sequencing analysis of 104 strains of the COVID-19 virus isolated from patients in different localities with symptom onset between the end of December 2019 and mid-February 2020 showed 99.9% homology, without significant mutation (Figure 1)."
"Laboratory, diagnostics and virology
The virus found to cause COVID-19 was initially isolated from a clinical sample on 7 January. It is notable that within weeks following the identification of the virus, a series of reliable and sensitive diagnostic tools were developed and deployed. On 16 January, the first RTPCR assays for COVID-19 were distributed to Hubei. Real-time PCR kits were distributed to all the provinces on 19 January and were provided to Hong Kong SAR and Macao SAR on 21 January. Information regarding viral sequences and PCR primers and probes was shared with WHO and the international community by China CDC on 12 January 2020. To facilitate product development and research on the new virus, COVID-19 virus sequences were uploaded to the GISAID Database by China.
By 23 February, there were 10 kits for detection of COVID-19 approved in China by the NMPA, including 6 RT-PCR kits, 1 isothermal amplification kit, 1 virus sequencing product and 2 colloidal gold antibody detection kits. Several other tests are entered in the emergency approval procedure. Currently, there are at least 6 local producers of PCR test kits approved by NMPA. Overall, producers have the capacity to produce and distribute as "many as 1,650,000 tests/week."
13.01.2020 : Christian Drosten (WHO) "We acknowledge ... release of another sequence (MN908947) ... We used known SARS- and SARS-related coronaviruses (bat viruses from our own studies
as well as literature sources) to generate a non-redundant alignment"
20.01.2020 : Christian Drosten (WHO) "virus isolates are unavailable," "We aimed to develop and deploy robust diagnostic methodology for use in public health laboratory settings without having virus material available."
21.07.2021 : United States CDC “Since no quantified virus isolates of the 2019-nCoV were available for CDC use at the time the test was developed and this study conducted, assays designed for detection of the 2019-nCoV RNA were tested with characterized stocks of in vitro transcribed full length RNA … spiked into a diluent consisting of a suspension of human A549 cells and viral transport medium (VTM) to mimic clinical specimen, ” CDC 2019-Novel Coronavirus (2019-nCoV) Real-Time RT-PCR Diagnostic Panel, page 41
20.02.2020 A Novel Coronavirus from Patients with Pneumonia in China, 2019
03.02.2020 A new coronavirus associated with human respiratory disease in China
28.02.2020 The Pathogenicity of SARS-CoV-2 in hACE2 Transgenic Mice
17.06.2020Severe Acute Respiratory Syndrome Coronavirus 2 from Patient with Coronavirus Disease, United States
"Therefore, we examined the capacity of SARS-CoV-2 to infect and replicate in several common primate and human cell lines, including human adenocarcinoma cells (A549), human liver cells (HUH7.0), and human embryonic kidney cells (HEK-293T), in addition to Vero E6 and Vero CCL81 cells. We also examined an available big brown bat kidney cell line (EFK3B) for SARS-CoV-2 replication capacity. Each cell line was inoculated at high multiplicity of infection and examined 24 h postinfection (Figure 3, panel A). No CPE was observed in any of the cell lines except in Vero cells"
10.01.2020 : WHO, " “The causal agent has not yet been identified or confirmed. The genetic sequence is not yet available. WHO DG is discussing with the Chinese Health Authorities to share the genetic sequence of the virus.”" WHO/Christian Drosten : “Laboratory testing of human suspected cases of novel coronavirus (nCoV) infection” despite “The causal agent has not yet been identified or confirmed,” refer to China/12.01.2019 and WHO/13.01.2020
11.01.2020 : WHO tweet “BREAKING: WHO has received the genetic sequences for the novel #coronavirus (2019-nCoV) from the Chinese authorities. We expect them to be made publicly available as soon as possible.” Refer to China/12.01.2020 and China/18.01.2020
13.01.2020 : WHO published the official protocol for coronavirus testing/RT-PCR
"...We used known SARS- and SARS-related coronaviruses (bat viruses from our own studies as well as literature sources) to generate a non-redundant alignment (excerpts shown in Annex). We designed candidate diagnostic RT-PCR assays before release of the first sequence of the Wuhan virus. Upon sequence release, three assays were selected based on their matching to the Wuhan virus as per inspection of the sequence alignment (Figures 1 and 2)...," Christian Drosten
14.01.2020
15.01.2020
16.01.2020
17.01.2020 : WHO's official protocol for detection of coronavirus
19.01.2020
20.01.2020 : Christian Drosten published the ‘Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR’
Background
The ongoing outbreak of the recently emerged novel coronavirus (2019-nCoV) poses a challenge for public health laboratories as virus isolates are unavailable while there is growing evidence that the outbreak is more widespread than initially thought, and international spread through travellers does already occur.
Aim
We aimed to develop and deploy robust diagnostic methodology for use in public health laboratory settings without having virus material available.
Methods
Here we present a validated diagnostic workflow for 2019-nCoV, its design relying on close genetic relatedness of 2019-nCoV with SARS coronavirus, making use of synthetic nucleic acid technology.
Results
The workflow reliably detects 2019-nCoV, and further discriminates 2019-nCoV from SARS-CoV. Through coordination between academic and public laboratories, we confirmed assay exclusivity based on 297 original clinical specimens containing a full spectrum of human respiratory viruses. Control material is made available through European Virus Archive – Global (EVAg), a European Union infrastructure project.
Conclusion
The present study demonstrates the enormous response capacity achieved through coordination of academic and public laboratories in national and European research networks.
21.01.2020
22.01.2020
23.01.2020> : Eurosurveillance published the ‘Detection of 2019 novel coronavirus (2019-nCoV) by real-time RT-PCR’ Corman-drosten Review Report: "The Drosten RT-PCR test is neither diagnostic for an infection with SARS-CoV-2 nor for a sickness or death from COVID-19. IT IS AN IMAGINARY VIRUS!"
24.01.2020
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09.02.2020
11.02.2020 : WHO announced that the disease caused by the novel coronavirus would be named COVID-19. Refer to 13.10.2015.
