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Drosten “SARS-COV-1” Paper (2003)


Christian Drosten, a German virologist who helped to create the problematic “SARS-COV-2” PCR test without ever actually seeing a “virus,” was also heavily involved in the original “SARS-COV-1” discovery process. In fact, Drosten was instrumental in setting up the molecular virology methods used today to claim the discovery of novel “viruses” based on genome sequences and online reference databases. He was the first to develop a “diagnostic” test for “SARS-COV-1” and coincidentally (or not) was also the first to develop a test for “SARS-COV-2” using nothing but social media reports. For a brief overview of Drosten’s role in the “SARS-COV-1” hoax, I have provided a few paragraphs from a Science article written in April 2020:

THE CORONAVIRUS CZAR

“Drosten’s coronavirus career effectively began on Saturday, 15 March 2003, when a 32-year-old doctor from Singapore named Leong Hoe Nam was taken off a plane in Frankfurt, Germany, and taken to the city’s university clinic. Leong had treated patients in Singapore before attending an infectious diseases course in New York City, and had developed symptoms consistent with an alarming new respiratory disease that was rapidly spreading in Asia. That same day, the World Health Organization (WHO) had christened the new disease “severe acute respiratory syndrome,” or SARS.

At the time, Drosten was building up a lab for molecular diagnostics at the Bernhard Nocht Institute for Tropical Medicine in Hamburg, Germany. The Frankfurt virologists sent Leong’s blood and other samples to Drosten, hoping he could help identify what was believed to be a new virus. But tests for everything from adenoviruses to paramyxoviruses came back negative.

About 1 week later, however, when Drosten was in Frankfurt to defend his Ph.D. thesis, the same virologists told him they had managed to grow the virus in a petri dish. Drosten realized this would allow him to use a new catch-all method he had developed for identifying unknown viruses, which amplified viral genetic material so it could be sequenced and checked against online databases. Drosten picked up a sample, then drove the 5 hours back to Hamburg in his old Opel and went straight to his lab. After a few days with little sleep, he had a small part of the new virus’ genome. The closest match was a cattle coronavirus that doesn’t infect people. “My first thought was, maybe it is some kind of contamination from the FCS,” the fetal calf serum used to grow cells in the lab, Drosten recalls.

But he and his colleague Stephan Günther quickly realized they were seeing a deadly new member of the family. “At the time, medical students learned hardly anything about coronaviruses,” Drosten says. The only two known to cause disease in humans, named OC43 and 229E, accounted for a small percentage of human colds every winter. This new virus was a very different beast. SARS killed 10% of the almost 8000 people it infected in nearly 30 countries before it was contained.

Researchers at the U.S. Centers for Disease Control and Prevention and at the University of Hong Kong realized the culprit was a coronavirus around the same time. But Drosten was the first to develop a diagnostic test, and he distributed the protocol freely on the internet. It earned him international recognition as well as the Federal Cross of Merit, an important German award.”

https://www.science.org/content/article/how-pandemic-made-virologist-unlikely-cult-figure

It’s interesting to note that Drosten originally thought that his “viral” genome for “SARS-COV-1” was actually contaminated from the Fetal Bovine Serum used in the cell culture to “isolate” his “virus.” It has been said that there is no way for the calf RNA to be freed from the cell culture. The FBS RNA is “co-isolated” with the cultured cells and the transcripts can be taken up by the cultured cells which can lead to false annotations and interfere with gene analysis:

Fetal Bovine Serum RNA Interferes with the Cell Culture derived Extracellular RNA

“Here, using RNA sequencing, we demonstrate that FBS contains a diverse repertoire of protein-coding and regulatory RNA species, including mRNA, miRNA, rRNA and snoRNA. The majority of them (>70%) are retained even after extended ultracentrifugation in the preparations of vesicle-depleted FBS (vdFBS) commonly utilized in the studies of extracellular vesicles (EV) and intercellular communication. FBS-associated RNA is co-isolated with cell-culture derived extracellular RNA (exRNA) and interferes with the downstream RNA analysis. Many evolutionally conserved FBS-derived RNA species can be falsely annotated as human or mouse transcripts. Notably, specific miRNAs abundant in FBS, such as miR-122, miR-451a and miR-1246, have been previously reported as enriched in cell-culture derived EVs, possibly due to the confounding effect of the FBS. Analysis of publically available exRNA datasets supports the notion of FBS contamination. Furthermore, FBS transcripts can be taken up by cultured cells and affect the results of highly sensitive gene expression profiling technologies. Therefore, precautions for experimental design are warranted to minimize the interference and misinterpretations caused by FBS-derived RNA.”

https://www.nature.com/articles/srep31175?fbclid=IwAR025YeZ64s3HyHaROFYnTSMbBmTHMQNRzmcDA8vBwQQs4Ao7oddPvqUk2M

Fetal Bovine Serum: blood taken from the heart of a still alive unborn baby calf of a slaughtered mother.

Maybe Drosten should have stuck with his first instinct?

In any case, here are some excerpts from his 2003 “SARS-COV-1” discovery paper which help to shed some light on the origins of the molecular virology scam being utilized today:

Identification of a Novel Coronavirus in Patients with Severe Acute Respiratory Syndrome

Methods

Clinical specimens from patients with SARS were searched for unknown viruses with the use of cell cultures and molecular techniques.

Results

A novel coronavirus was identified in patients with SARS. The virus was isolated in cell culture, and a sequence 300 nucleotides in length was obtained by a polymerase-chain-reaction (PCR)–based random-amplification procedure. Genetic characterization indicated that the virus is only distantly related to known coronaviruses (identical in 50 to 60 percent of the nucleotide sequence). On the basis of the obtained sequence, conventional and real-time PCR assays for specific and sensitive detection of the novel virus were established. Virus was detected in a variety of clinical specimens from patients with SARS but not in controls. High concentrations of viral RNA of up to 100 million molecules per milliliter were found in sputum. Viral RNA was also detected at extremely low concentrations in plasma during the acute phase and in feces during the late convalescent phase. Infected patients showed seroconversion on the Vero cells in which the virus was isolated.

Conclusions

The novel coronavirus might have a role in causing SARS.”

Microbiologic testing for common
pathogens

“A large number of tests for known respiratory pathogens were performed with specimens from all three patients in Frankfurt. The test results were negative, except as follows.

Paramyxovirus-like particles were seen in throat swabs and sputum samples from the index patient by electron microscopy. The particles were scarce. However, several PCR tests specific for virus species of the family Paramyxoviridae were negative (including tests for human metapneumovirus), as were PCR assays based on primers designed to react broadly with all members of that family.

C. pneumoniae was not detected by PCR or antigen ELISA in sputum of the index patient from day 9. However, on day 11, electron microscopy of cells in a bronchoalveolar-lavage specimen from the index patient showed a severe intracellular bacterial infection, and the bronchoalveolar-lavage cells reacted in
immunofluorescence analyses with a monoclonal antibody directed against C. pneumoniae. Consistent with this finding, there was an increase by a factor of four in the C. pneumoniae IgA titer in the index patient between day 10 and day 13.

Isolation and characterization of a novel coronavirus

After six days of incubation (on March 21), a cytopathic effect was seen on Vero-cell cultures inoculated with sputum obtained from the index patient on day 7. Twenty-four hours after a single passage, nucleic acids were purified from the supernatant. Random amplification was performed with 15 different PCRs under low-stringency conditions. We had previously shown that this method is able to detect unknown pathogens growing in cell culture (unpublished data). To detect RNA viruses, an initial reverse-transcription step was included.

About 20 distinct DNA fragments were obtained and sequenced. The resulting sequences were subjected to BLAST data-base searches. Most of the fragments matched human chromosome sequences, indicating that genetic material of the cultured cells had been amplified (Vero cells are derived from monkeys). Three of the fragments did not match any nucleotide sequence in the data base. However, when a translated BLAST search was performed (comparison of the amino acid translation in all six possible reading frames with the data base), these fragments showed homology to coronavirus amino acid sequences, indicating that a coronavirus had been isolated. Two of the fragments were 300 nucleotides in length and identical in sequence, and the third fragment was 90 nucleotides in length (sequences BNI-1 and BNI-2, respectively, as reported on the Web site of the WHO network on March 25) (Fig. 1A). Detailed sequence analysis revealed that both fragments were located in the open reading frame 1b of coronaviruses and did not overlap with a 400-nucleotide coronavirus fragment identified by colleagues at the Centers for Disease Control and Prevention (CDC) (sequence CDC, reported on the Web site of the WHO network on March 24) (Fig. 1A).

There were sequences of six coronaviruses of all phylogenetic lineages (groups 1, 2, and 3) in the sequence data base that overlapped with sequences BNI-1 and BNI-2. All known sequences were only distantly related to the new sequences (Fig. 1B). The BNI-1 nucleotide sequence diverged from the known sequences by between 39 percent (bovine coronavirus) and 46 percent (porcine epidemic diarrhea virus). Such distances are typically found between different genetic lineages of coronaviruses.7
A stable phylogeny was obtained with the BNI-1 fragment, indicating that the isolated coronavirus segregates between genetic groups 2 and 3 (Fig. 1C).”

The established PCR assays have also been used to test respiratory samples from German patients with symptoms and a travel history compatible with SARS. So far, 67 samples from 55 patients have been tested. One patient fulfilling the WHO criteria for probable SARS was coronavirus-positive on PCR. PCR protocols, as well as positive control material, have been made available to laboratories worldwide.”

Discussion

The principal finding of the study is the identification of a novel coronavirus in patients with SARS. It appears that patients with SARS are acutely infected with this virus, since they have virus-specific IgG seroconversion. The high rate of positivity among patients with probable cases during an outbreak of SARS in Hanoi, in conjunction with the complete negativity among all healthy contacts of patients affected by the same outbreak, provides evidence of an association between the disease and the presence of this novel virus. The involvement of a coronavirus in a respiratory disease would not be without precedent: the two human coronaviruses are known to cause mild respiratory illness.9 One should bear in mind, however, that in the past, viruses have been initially isolated from patients with a specific disease but subsequent investigations revealed no actual association at all.10,11 Thus, larger studies with appropriate control groups are needed to verify or eliminate our hypothesis about the cause of SARS. The assays that have been established provide an excellent tool for such studies. It should also be taken into account that antigen that was present in primary cultures was used to detect the antibody response, and it remains to be firmly established that this response is indeed directed against the novel coronavirus rather than against an unknown agent that might have been isolated simultaneously. This possibility could be tested with the use of plaque-purified virus or recombinant proteins as antigen.

By testing for a broad range of known pathogens, we also obtained evidence for infection with paramyxoviruses and C. pneumoniae. Paramyxoviruses —in particular, human metapneumovirus, which was previously implicated in SARS— could be largely ruled out by further investigation. Infection with chlamydia was confirmed in several assays. However, chlamydia was not found in other patients with SARS.3 Hence, it remains unclear whether these pathogens have a role as causative factors or cofactors in SARS.

doi: 10.1056/NEJMoa030747

Presenting “SARS-COV-1!!!” That’s convincing, right?

In Summary:

  • The aim of study was to detect unknown “viruses” by way of indirect methods such as cell culture and molecular techniques
  • Drosten et al claim the “virus” was “isolated” in cell culture which is the exact opposite of isolation due to the adding/mixing of various foreign animal DNA, chemicals, nutrients, antibiotics, etc. with the sample
  • They utilized a random-amplification PCR procedure to detect sequence of the “virus” in cell culture fluids
  • The “virus” was considered to be distantly related to known “Coronaviruses” (only 50-60% identical in nucleotide sequences)
  • They created their own PCR tests to detect parts of this sequence in “SARS” patients
  • Drosten determines that the “virus” MIGHT HAVE A ROLE in causing “SARS”
  • Paramyxovirus-like particles and C. Pneumoniae were also found in samples
  • CPE was seen after 7 days of incubation
  • Random amplification of culture fluid by PCR was performed to detect a “novel virus”
  • They state that this method was shown to be able to detect unknown pathogens in UNPUBLISHED RESEARCH

(Quick side note. This is from a study done in 2019, sixteen years after Drosten’s paper stated they could detect unknown pathogens by PCR. This paper, along with examples from previous studies, state this is impossible. From the April 2019 study:

A novel method for the capture-based purification of whole viral native RNA genomes

“However, extraction of viral RNA directly from culture often yields viral RNA with high host RNA background (Marston et al. 2013).”

“Similarly, Cowan et al. (2005) noted that PCR-based enrichment techniques where a priori knowledge of target sequences is required for PCR primer design render the enrichment strategies ineffective in the characterization of novel viruses. It follows that the same sequence ambiguity in viral genomes would pose a problem for using customized sequencing adapters during direct RNA sequencing.

From the abovementioned problems, we identified a need for an alternative technique that isolates viral RNA molecules from host RNA background without PCR amplification or cDNA synthesis.

https://amb-express.springeropen.com/articles/10.1186/s13568-019-0772-y

Maybe this is why Drosten’s data proving his method went UNPUBLISHED…but I digress…)

  • The resulting sequences were subjected to BLAST data-base searches for comparison to known sequences
  • Three short fragments (two 300 nucleotides and one 90 nucleotides) showed homology to “coronavirus” amino acid sequences, which indicated to them that a “coronavirus” had been “isolated”
  • All known sequences were only distantly related to the new fragments
  • The BNI-1 nucleotide sequence diverged from the known sequences by between 39 percent (bovine “coronavirus”) and 46 percent (porcine epidemic diarrhea “virus”)
  • The Drosten PCR test was used on 67 samples from 55 German cases of probable “SARS” and only one patient fulfilling the WHO criteria for probable “SARS” was coronavirus-positive on PCR
  • Drosten et al assume patients with SARS are infected due to “virus-specific” seroconversion which is impossible to determine without a purified/isolated “virus”
  • They claim that the high rate of positivity among patients with probable cases during an outbreak of “SARS” in Hanoi, in conjunction with the complete negativity among all healthy contacts of patients affected by the same outbreak, provides evidence of an association between the disease and the presence of this novel “virus” yet this obviously shows it to not be highly transmissable/contagious
  • They admit that “viruses” have been “isolated” in the past which were believed to be the cause of disease and it turned out that there was no association with the disease
  • They admit larger studies with appropriate controls need to be done in order to verify or eliminate their own “SARS” hypothesis
  • They admit that there needs to be confirmation that the antibody response is due to this new “coronavirus” rather than some unknown agent which may have been “isolated” along with it
  • Human metapneumovirus was previously implicated in “SARS”
  • Since Paramyxoviruses, C. Pneumoniae, and Chlamydia was also “isolated” from “SARS” patients, it remains unclear whether these also play a role in “SARS”

Note in this paper that once again there was no properly purified/isolated “virus” taken directly from a sick patient and proven pathogenic. There were no accompanying EM images as they used an indirect PCR method to sequence an unknown “virus.” There were only three small sequence fragments of unknown origin that were distantly related to known “Coronaviruses,” specifically from cows and pigs. Drosten himself initially believed his “isolate” was contaminated by the fetal bovine serum used to culture the “virus.” Was this possibility thoroughly investigated? Not according to this paper. On top of this, they admit that larger studies with appropriate controls needed to be performed to either confirm or deny their “SARS” hypothesis. In other words, the only evidence Drosten et al provide that a new “virus” exists at all is a hypothesis which comes from random letters in a computer database and a PCR test unsuitable to discover novel “viruses.”

Full article here: https://viroliegy.com/2021/10/15/drosten-sars-cov-1-paper-2003/

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This entry was posted on October 16, 2021 by in Coronavirus, Fraud, Globalism, Health and tagged , , , , .

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