Cycle Thresholds RT-PCR
Open letter regarding Public Health Strategies.
This is an open letter directed to all people who may have an influence on matters regarding public health strategies, specifically on measures relating to SARS-Cov2.
To all local councillors, politicians (MPs, MSPs, MEPs) of all political parties or independents, doctors, health professionals, academics, and concerned citizens ~
There is something of importance, which I would like to draw to your attention. I do this in the hope that you will consider the significance, inspire you to do some research of this topic, and debate with others. This topic affects you, your family, everyone in this country, and citizens of other countries. Science is a journey, it does not begin with the destination.
We have heard many times catchphrases such as 'we are all in this together' or 'we are following the science'. However, most people now realise there are anomalies with both of these statements. I am a great believer in the power of knowledge; I also believe that the more knowledge people have of a particular thing, then the greater the power those people will have in influencing how that particular thing affects them, directly or indirectly.
The subject matter is complicated but that should not prevent it being openly debated, if we are truly all in this together and following the science. The topic I am asking you to take time to read about, get a basic understanding of, and debate, is that of...
'Cycle Thresholds' used in the RT-PCR technique, which you may be aware of as being used to 'test' for SARS-Cov2.
To be clear from the outset -
There is nothing being recommended or proposed here, except the acquisition of knowledge and the need to debate. The following web links include numerous news articles or opinions or science papers which may or may not be peer-reviewed and therefore should not be taken as established facts.
Any mention of products does not imply endorsement.
The sole purpose of gathering these links together is simply to highlight the topic urgently requiring debate. This is particularly important for two reasons related to testing:
Firstly, it may lead to making it easier to focus on helping those who are most at risk and thereby reducing the risk of spreading a virus.
Secondly, it may lead to removing a lot of the false positives and filter those people who are most likely to be non-infective and thereby allowing more freedom for their daily activities, which in turn, would also benefit in freeing up the resources (human and monetary) needed for those who are at risk of illness and/or in need of treatment or financial help.
In order to help you in this learning process, I have provided multiple links to various websites below, beginning with some general background, explanation of terms etc., and then leading to some studies and news articles that I believe show that the PCR technique (a research tool) may not be currently used to its full advantage.
To assist people that perhaps do not have much (nor any) experience in reading scientific matters, which can appear daunting, I will extract the most important details of each link and place them in quotation marks. However, please try to read the actual papers and/or media articles the links refer to, especially any conclusions.
Please try, at least, to get to the bottom of this page, especially to the link regarding the efficacy of Vitamin D3 as a preventative measure!
If reading about PCR is really too difficult for you, then you may wish to skip down the page to the paragraph starting "***Regarding public health strategies and INFECTIVITY -"
The technique known as Polymerase Chain Reaction (PCR) used as a research technique was invented by the late Dr Karey Banks Mullis 1944~2019. See KareyMullis.com
Dr Mullis apparently had an epiphany whilst driving - "I very quickly brought the Honda to a stop near the roads edge... After ten cycles I would have a thousand... Thirty cycles would be somewhere around a billion. The product would overwhelm anything that was unintended because it would be self catalytic, and only the site of interest would bind the necessary two oligos together in their little reproductive dance.
I didn't sleep that night. The next morning I bought two bottles of Navarro Vineyards Pinot Noir, and by mid afternoon had settled into a fitful sleep. There were diagrams of PCR reactions on every surface that would take pencil or crayon in my cabin. I woke up in a new world." Source ~ KareyMullis.com/pcr
Introduction to PCR
The polymerase chain reaction (PCR) is a relatively simple technique that amplifies a DNA template to produce specific DNA fragments in vitro. Click here for source ~ Promega.co.uk
However, SARS-cov2 is thought to be an RNA virus. To analyse RNA, an additional step is required beforehand, this is called Reverse Transcription (RT). Then the process is referred to as RT-PCR. It should be noted that there is a version called real time RT-PCR, however RT does not stand for real time... "For optimal results, the RNA template, whether a total RNA preparation, an mRNA population or a synthesized RNA transcript, should be DNA-free to avoid amplification of contaminating DNA. The most commonly used DNA polymerases for PCR have no reverse transcriptase activity under standard reaction conditions, and thus, amplification products will be generated only if the template contains trace amounts of DNA with similar sequences." Click for source ~ Promega.co.uk
"General Considerations for RT-PCR
For RT-PCR, successful reverse transcription depends on RNA integrity and purity. Procedures for creating and maintaining a ribonuclease-free (RNase-free) environment to minimize RNA degradation are described in Blumberg, 1987." Click for source ~ Promega.co.uk
|
"Real-time PCR, also called quantitative PCR or qPCR, can provide a simple and elegant method for determining the amount of a target sequence or gene that is present in a sample. Its very simplicity can sometimes lead to problems by overlooking some of the critical factors that make it work. This review will highlight these factors that must be considered when setting up and evaluating a real-time PCR reaction."
|
***Regarding public health strategies and INFECTIVITY - Why are people not talking about Cycle Threshold(s)? The RT-PCR technique (a research tool) is open to interpretation; perhaps it should not be used as a binary test (Yes/No) when it comes to infectivity because of variations in viral load or non-infective fragments of RNA. The cycle threshold (CT) may indicate this (through correlation, see links below) but is not being mentioned. To directly compare results from different test centres (globally) the CT would have to be identical. I can find no evidence that this is indeed the case. A binary result (Y/N) does not provide useful information regarding infectivity. EXTRACT "this may not be optimal because it assumes a positive PCR test is intended to mean infectivity. Closer examination of what the test results mean clinically, particularly when results are from RNA quantities near the lower limit of detection of the assay, could help guide clinical and public health strategies." Source ~ Click here for article
~ To Interpret the SARS-CoV-2 Test, Consider the Cycle Threshold Value
|
The cycle threshold of detection of what is known as SARS-cov2 has massive implications for public health strategies... "Correlation between successful isolation of virus in cell culture and Ct value of quantitative RT-PCR targeting E gene suggests that patients with Ct above 33–34 using our RT-PCR system are not contagious and thus can be discharged from hospital care or strict confinement for non-hospitalized patients." ~
Click for source ~ Viral RNA load as determined by cell culture as a management tool for discharge of SARS-CoV-2 patients from infectious disease wards
|
Similarly, in a quote from The University of Louisville - "Our current approach is to allow to return to work healthcare workers with persistently positive RT-PCR if the Ct (cycle threshold) values are greater than 35." Click for Source ~ The University of Louisville
|
Surprisingly, NHS England is using a 'cycle threshold' of 45 cycles as an indicator of infectivity. At present, I have no information on the CT value being used in Scotland. I would have thought somewhere between 25 - 30 cycles would be more realistic from what I've been reading, especially on a second test. Then the focus could be on those who need treatment and/or quarantine. Cycle Thresholds mentioned just after 12 minutes, speaker Professor Carl Heneghan... Source ~ Westminster gov policy YouTube
Also, see p.16 of this document - Royal College of Pathologists
|
Why does the cycle threshold cut-off matter?
"RT-PCR uses an enzyme called reverse transcriptase to change a specific piece of RNA into a matching piece of DNA. The PCR then amplifies the DNA exponentially, by doubling the number of molecules time and again. A fluorescent signal can be attached to the copies of the DNA, and a test is considered positive when the fluorescent signal is amplified sufficiently to be detectable.
The cycle threshold (referred to as the Ct value) is the number of amplification cycles required for the fluorescent signal to cross a certain threshold. This allows very small samples of RNA to be amplified and detected... The lower the cycle threshold level the greater the amount of RNA (genetic material) there is in the sample. The higher the cycle number, the less RNA there is in the sample.
What does this mean?
This detection problem is ubiquitous for RNA viruses detection. SARS-CoV, MERS, Influenza Ebola and Zika viral RNA can be detected long after the disappearance of the infectious virus.
The immune system works to neutralise the virus and prevent further infection. Whilst an infectious stage may last a week or so, because inactivated RNA degrades slowly over time it may still be detected many weeks after infectiousness has dissipated."
Click for source ~ Are you infectious if you have a positive PCR test result for COVID-19?
|
Are public health policies in different countries or states based on an arbitrary number of cycle thresholds in RT-PCR?
NHS England uses 45 cycles to judge infectivity (Scotland?). USA possibly uses different CTs for different states.
"Up to 90 percent of people tested for COVID-19 in Massachusetts, New York and Nevada in July carried barely any traces of the virus and it could be because today's tests are 'too sensitive', experts say... Health experts say PCR testing - the most widely used diagnostic test for COVID-19 in the US - are too sensitive and need to be adjusted to rule out people who have insignificant amounts of the virus in their systems because they're likely not contagious.
Today the PCR test, which provides a yes or no answer if a patient is infected, doesn't say how much of the virus a patient has in their body...
"New York's state lab Wadsworth analyzed cycle thresholds values in already processed COVID-19 PCR tests and found in July that 794 positive tests were based on a threshold of 40 cycles...
Researchers say the solution is even more widespread use of rapid tests with an adjusted threshold to hone in on the most infectious people with COVID-19. According to Dr Michael Mina, an epidemiologist at the Harvard T.H. Chan School of Public Health, said to the Times, 'We’ve been using one type of data for everything, and that is just plus or minus – that’s all. We’re using that for clinical diagnostics, for public health, for policy decision-making... It’s really irresponsible, I think, to forgo the recognition that this is a quantitative issue'. Experts say a reasonable cutoff for the virus would be 30 or 35 cycles, according to Juliet Morrison, a virologist at the University of California, Riverside.
Dr Mina said he would set the cut-off at 30.
With a cutoff of 35, about half of those tests would no longer qualify as positive. About 70 percent would no longer be judged positive if the cycles were limited to 30... Massachusetts, from 85 to 90 percent of people who tested positive in July with a cycle threshold of 40 would have been considered negative if the threshold were 30 cycles, Dr Mina said - 'I would say that none of those people should be contact-traced, not one,'."
Click for source ~ Experts: US COVID-19 positivity rate high due to 'too sensitive' tests
|
In an article in the 'Infectious Disease Advisor' written by Sweta Gupta - "Correlation between successful isolation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in cell culture and cycle threshold (Ct) value of quantitative reverse transcription polymerase chain reaction (RT-PCR) targeting E gene suggests that patients with coronavirus disease 2019 (COVID-19) with Ct above 33 to 34 are not contagious and can be discharged from hospital care or strict confinement, according to a brief report published in the European Journal of Clinical Microbiology & Infectious Diseases."
Click for source ~ Strong Inverse Correlation Between SARS-CoV-2 Infectivity and Cycle Threshold Value
|
In this (not yet peer reviewed) article 'Antigen testing detects SARS-Cov2 better than PCR', Dr Sanchari Dutta PhD writes - "A team of scientists from the United States demonstrates that rapid antigen tests for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are more effective in determining the actual infection status in patients with coronavirus disease 2019 (COVID-19) than real-time polymerase chain reaction (RT PCR)-based tests..."
Important notice - "medRxiv publishes preliminary scientific reports that are not peer-reviewed and, therefore, should not be regarded as conclusive, guide clinical practice/health-related behaviour, or treated as established information."
Click for source ~ News-Medical.net
|
We have been hearing very little about false positives in the news.
"Why are false positives a problem? DHSC figures [3] show that 100,664 tests were carried out on 31 May 2020 (Pillar 1 and 2 RT-PCR tests). 1,570 of those tests were positive for SARS-CoV-2 (1.6%). The majority of people tested on that day did not have SARS-CoV-2 (98.4% of tests are negative). When only a small proportion of people being tested have the virus, the operational false positive rate becomes very important. Clearly the false positive rate cannot exceed 1.6% on that day, and is likely to be much lower. If the operational false positive rate was 0.4%, 400 of the 1,570 positive tests would be false positives. That would represent 400 people being isolated when they are well, and much wasted effort in contact tracing. It is possible that a proportion of infections that we currently view as asymptomatic may in fact be due to these false positives. Unless we understand the operational false positive rate of the UK’s RT-PCR testing system we risk overestimating the COVID-19 incidence, the demand on track and trace, and the extent of asymptomatic infection."... "The UK operational false positive rate is unknown. There are no published studies on the operational false positive rate of any national COVID-19 testing programme." Please note, there appears to be NO mention of the cycle thresholds used in this document.
Click for source ~ Gov.uk web page
Clearly, there is some awareness of false positives and the reasons for them. Here is a short extract from The Lancet (Respiratory Medicine) - "When low pretest probability exists, positive results should be interpreted with caution and a second specimen tested for confirmation. Notably, current policies in the UK and globally do not include special provisions for those who test positive despite being asymptomatic and having laboratory confirmed COVID-19 in the past (by RT-PCR swab test or antibodies). Prolonged viral RNA shedding, which is known to last for weeks after recovery, can be a potential reason for positive swab tests in those previously exposed to SARS-CoV-2. However, importantly, no data suggests that detection of low levels of viral RNA by RT-PCR equates with infectivity unless infectious virus particles have been confirmed with laboratory culture-based methods." Click for source ~ TheLancet.com
|
Where did the use of RT-PCR (a research tool) emanate as a 'diagnostic test' for SARS-cov2?
Extract -
"Christian Heinrich Maria Drosten (German: born 1972 is a German virologist whose research focus is on novel viruses (emergent viruses). During the COVID-19 pandemic, Drosten came to national prominence as an expert on the implications and actions required to combat the outbreak in Germany." Click for Source ~ Christian Drosten Wikipedia.
From the above source -
"On 23 January 2020, Drosten, along with other virologists in Europe and Hong Kong, published a workflow of a real-time PCR (RT-PCR) diagnostic test, which was quickly accepted by the World Health Organization (WHO) who sent test kits to affected regions...
In March 2020, he was appointed to the European Commission's advisory panel on COVID-19, co-chaired by Ursula von der Leyen and Stella Kyriakides."
See 'Diagnostic detection of 2019-nCoV by real-time RT-PCR' source ~ WHO Protocol v2
Where does the 45 cycle threshold used in RT-PCR emanate?
According to the following source, the 45 cycle threshold was advocated by Christian Drosten et al. "Thermal cycling was performed at 55 °C for 10 min for reverse transcription, followed by 95 °C for 3 min and then 45 cycles of 95 °C for 15 s, 58 °C for 30 s... Authors' Contributions... CD: Planned experiments, conceptualised the laboratory work, conceptualised the overall study, wrote the manuscript draft." Source ~
Eurosurveillance - Detection of 2019 novel coronavirus... by real time RT-PCR
|
...
Has the RT-PCR technique been used in a similar way before?
HIV - Yes. Extract - "Kary Mullis, who won the Nobel Prize in Science for inventing the PCR, is thoroughly convinced that HIV is not the cause of "AIDS". With regard to the viral load tests, which attempt to use PCR for counting viruses, Mullis has stated: "Quantitative PCR is an oxymoron." PCR is intended to identify substances qualitatively, but by its very nature is unsuited for estimating numbers. Although there is a common misimpression that the 'viral load tests' actually count the number of viruses in the blood, these tests cannot detect free, infectious viruses at all; they can only detect proteins that are believed, in some cases wrongly, to be unique to HIV. The tests can detect genetic sequences of viruses, but not viruses themselves." Click for source virusmyth.org
An example of a false positive HIV 'case' due to inappropriate use of HIV PCR 'case' -
"Our urge to confirm the cause of acute encephalopathy rather than accept a diagnosis of exclusion resulted in inappropriate use of HIV-1 PCR."
Click for source False positive HIV diagnosis.
Further evidence that viral load may be inversely correlated to the cycle threshold may be seen in studies involving young children and HIV. For example - "HIV-1 viral load (VL) has been found to be an independent predictor for disease progression among untreated HIV-infected children. However, qualitative polymerase chain reaction (PCR) assays are routinely used for early infant diagnosis (EID)...
Conclusion: EID Ct values at birth predict VL {viral load}..."
Click for source Early Infant Diagnosis HIV-1 PCR cycle-threshold predicts.
A video comment from Karey Mullis, the inventor of PCR, regarding the 'interpretation' or 'misinterpretation' of the PCR technique pertaining to HIV detection. ~ YouTube video.
SARS (Severe Acute Respiratory Syndrome) - YES.
"There have been 2 self-limiting SARS outbreaks, which resulted in a highly contagious and potentially life-threatening form of pneumonia. Both happened between 2002 and 2004.
Since 2004, there have not been any known cases of SARS reported anywhere in the world." Source ~ NHS UK Conditions SARS. See also Wikipedia ~ Wikipedia SARS
The scientist Christian Drosten was "the first to develop a diagnostic test, and he distributed the protocol freely on the internet." Click for source ~ Sciencemag.org
The WHO incorporated PCR as a diagnosis. Click for source ~ WHO LabMethods
MERS (Middle East Respiratory Syndrome) - YES.
According to the NHS - "Middle East respiratory syndrome coronavirus (also known as MERS or MERS-CoV) is a rare but severe respiratory illness. It can start with a fever and cough, which can develop into pneumonia and breathing difficulties.
MERS was first identified in 2012 in the Middle East and is most common in that region.
The risk of infection with MERS to people in the UK is very low." Source ~ NHS MERS
See also source update from the WHO ~ the WHO MERS Disease outbreak update
Also see WHO's document revised 2018 ~ Laboratory Testing for Middle East Respiratory Syndrome Coronavirus
According to ScienceMag "Scientists discovered two new coronaviruses in the years after the SARS outbreak, both of which caused the common cold. Then in 2012, researchers isolated a new coronavirus that spelled greater danger. It came from a 60-year-old man in Saudi Arabia who had developed pneumonia.Intrigued, Christian Drosten geared his research to the new agent, which was soon called Middle East respiratory syndrome (MERS) virus. In 2013, he reported on a wealthy 73-year-old patient from Abu Dhabi, United Arab Emirates, who was treated for MERS in Germany and died. Relatives said the patient had cared for a sick racing camel before falling ill—the first sign that camels might be involved.
Saudi Arabia, which had the most MERS cases and a multimillion-dollar camel racing industry, initially scoffed at the link. “We don’t think camels are involved,” then–Deputy Minister of Health Ziad Memish said. But work from Drosten’s group and others soon confirmed the suspicion. Memish and Drosten teamed up to study the new disease, and Drosten’s Bonn lab became a leading MERS hub. It developed a test to detect the virus’ RNA and then an antibody assay that helped show the virus had likely been infecting people in the region for decades.
The research yielded some unexpected insights. While looking for coronaviruses in camels, the scientists found pathogens closely related to 229E, one of the common cold coronaviruses, suggesting that virus, too, originated in camels. It was a warning sign, Drosten said at the time, that MERS could follow the same course as SARS, which had originated in bats, and evolve to become a true human disease. Animal coronaviruses, it seemed, posed a particular threat of sparking a pandemic."
Click for source ~ ScienceMag.org
According to the Lancet, Professor Christian Drosten et al conducted an investigation of a patient with MERS using RT-PCR. Click for source ~ The Lancet MERS coronavirus
Whooping Cough (Pertussis) - YES.
A Whooping cough 'epidemic' that was based on false results - According to The New York Times, in an article dated Jan 22nd 2007, "Dr. Brooke Herndon, an internist at Dartmouth-Hitchcock Medical Center, could not stop coughing. For two weeks starting in mid-April last year, she coughed, seemingly nonstop, followed by another week when she coughed sporadically, annoying, she said, everyone who worked with her.
Before long, Dr. Kathryn Kirkland, an infectious disease specialist at Dartmouth, had a chilling thought: Could she be seeing the start of a whooping cough epidemic? By late April, other health care workers at the hospital were coughing, and severe, intractable coughing is a whooping cough hallmark. And if it was whooping cough, the epidemic had to be contained immediately because the disease could be deadly to babies in the hospital and could lead to pneumonia in the frail and vulnerable adult patients there...
Nearly 1,000 health care workers at the hospital in Lebanon, N.H., were given a preliminary test and furloughed from work until their results were in; 142 people, including Dr. Herndon, were told they appeared to have the disease; and thousands were given antibiotics and a vaccine for protection. Hospital beds were taken out of commission, including some in intensive care.
Then, about eight months later, health care workers were dumbfounded to receive an e-mail message from the hospital administration informing them that the whole thing was a false alarm. Not a single case of whooping cough was confirmed with the definitive test, growing the bacterium, Bordetella pertussis, in the laboratory.
Instead, it appears the health care workers probably were afflicted with ordinary respiratory diseases like the common cold. Now, as they look back on the episode, epidemiologists and infectious disease specialists say the problem was that they placed too much faith in a quick and highly sensitive molecular test that led them astray... At Dartmouth the decision was to use a test, PCR, for polymerase chain reaction. It is a molecular test that, until recently, was confined to molecular biology laboratories... Anyone who had a cough got a PCR test, and so did anyone with a runny nose who worked with high-risk patients like infants.
“That’s how we ended up with 134 suspect cases,” Dr. Kirkland said. And that, she added, "was why 1,445 health care workers ended up taking antibiotics and 4,524 health care workers at the hospital, or 72 percent of all the health care workers there, were immunized against whooping cough in a matter of days.
If we had stopped there, I think we all would have agreed that we had had an outbreak of pertussis and that we had controlled it,” Dr. Kirkland said...
Dr. Cathy A. Petti, an infectious disease specialist at the University of Utah, said the story had one clear lesson.
“The big message is that every lab is vulnerable to having false positives,” Dr. Petti said. “No single test result is absolute and that is even more important with a test result based on PCR.”
Click for source ~ The New York Times nytimes.com
|
In a review by Jefferson et al, they said that "A very small proportion of people re-testing positive after hospital discharge or with high Ct are likely to be infectious." They concluded by saying - "Prospective routine testing of reference and culture specimens are necessary for each country involved in the pandemic to establish the usefulness and reliability of PCR for Covid-19 and its relation to patient factors. Infectivity is related to the date of onset of symptoms and cycle threshold level. A binary Yes / No approach to the interpretation [sic] RT-PCR unvalidated against viral culture will result in false positives with possible segregation of large numbers of people who are no longer infectious and hence not a threat to public health." - Bold added here for emphasis. Click for source ~ Viral cultures for COVID-19 infectivity assessment. Systematic review.
|
Has the SARS-cov2 virus been isolated and purified?
On the GlobalResearch website, they investigated whether the SARS-cov2 virus RNA had been isolated and purified - "because the PCR tests are calibrated for gene sequences (in this case RNA sequences because SARS-CoV-2 is believed to be a RNA virus), we have to know that these gene snippets are part of the looked-for virus. And to know that, correct isolation and purification of the presumed virus has to be executed.
Hence, we have asked the science teams of the relevant papers which are referred to in the context of SARS-CoV-2 for proof whether the electron-microscopic shots depicted in their in vitro experiments show purified viruses.
But not a single team could answer that question with “yes” — and NB., nobody said purification was not a necessary step. We only got answers like “No, we did not obtain an electron micrograph showing the degree of purification” (see below).
We asked several study authors “Do your electron micrographs show the purified virus?”, they gave the following responses:
Study 1: Leo L. M. Poon; Malik Peiris. “Emergence of a novel human coronavirus threatening human health” Nature Medicine, March 2020
Replying Author: Malik Peiris
Date: May 12, 2020
Answer: “The image is the virus budding from an infected cell. It is not purified virus.”
Study 2: Myung-Guk Han et al. “Identification of Coronavirus Isolated from a Patient in Korea with COVID-19”, Osong Public Health and Research Perspectives, February 2020
Replying Author: Myung-Guk Han
Date: May 6, 2020
Answer: “We could not estimate the degree of purification because we do not purify and concentrate the virus cultured in cells.”
Study 3: Wan Beom Park et al. “Virus Isolation from the First Patient with SARS-CoV-2 in Korea”, Journal of Korean Medical Science, February 24, 2020
Replying Author: Wan Beom Park
Date: March 19, 2020
Answer: “We did not obtain an electron micrograph showing the degree of purification.”
Study 4: Na Zhu et al., “A Novel Coronavirus from Patients with Pneumonia in China”, 2019, New England Journal of Medicine, February 20, 2020
Replying Author: Wenjie Tan
Date: March 18, 2020
Answer: “[We show] an image of sedimented virus particles, not purified ones.”
"Regarding the mentioned papers it is clear that what is shown in the electron micrographs (EMs) is the end result of the experiment, meaning there is no other result that they could have made EMs from.
That is to say, if the authors of these studies concede that their published EMs do not show purified particles, then they definitely do not possess purified particles claimed to be viral. (In this context, it has to be remarked that some researchers use the term “isolation” in their papers, but the procedures described therein do not represent a proper isolation (purification) process. Consequently, in this context the term “isolation” is misused)."
Later in the article they say "It should not go unmentioned that we finally got the Charité – the employer of Christian Drosten, Germany’s most influential virologist in respect of COVID-19, advisor to the German government and co-developer of the PCR test which was the first to be “accepted” (not validated!) by the WHO worldwide – to answer questions on the topic.
But we didn’t get answers until June 18, 2020, after months of non-response. In the end, we achieved it only with the help of Berlin lawyer Viviane Fischer.
Regarding our question “Has the Charité convinced itself that appropriate particle purification was carried out?,” the Charité concedes that they didn’t use purified particles.
And although they claim “virologists at the Charité are sure that they are testing for the virus,” in their paper (Corman et al.) they state:
RNA was extracted from clinical samples with the MagNA Pure 96 system (Roche, Penzberg, Germany) and from cell culture supernatants with the viral RNA mini kit (QIAGEN, Hilden, Germany),”
Which means they just assumed the RNA was viral."
Click for source ~ Globalresearch covid19 PCR tests-
|
The RT-PCR technique may be mentioned by WHO as NAATs - "Standard confirmation of acute SARS-CoV-2 infections is based on the detection of unique viral sequences by nucleic acid amplification tests (NAATs), such as real-time reverse-transcription polymerase chain reaction (rRT-PCR). The assays’ targets include regions on the E, RdRP, N and S genes."
Download pdf here at source ~ WHO Publications Diagnostic Testing
Two letters make a difference; suitable or unsuitable ~ WHO Diagnostic testing for SARS-CoV-2: interim guidance, 11 September 2020
|
There is a fundamental need to standardise information reported to science journals with regard to RT-PCR - Stephen Bustin poses the question -
Why the need for qPCR publication guidelines?--The case for MIQE
|
Update 16th July 2020 ~ Click (video relevant at 3' 50") - Dr Anthony Fauci said "if you get a Cycle Threshold of 35 or more, the chances of it being replication competent are miniscule."
Update October 2020 ~ Click - UK Gov publishes info "RT-PCR detects presence of viral genetic material in a sample but is not able to distinguish whether infectious virus is present."
Update November 2020 ~ Click- Portuguese Court Rules PCR Tests as Unreliable
Update 27th November 2020 Click ~ CORMAN-DROSTEN REVIEW REPORT CURATED BY AN INTERNATIONAL CONSORTIUM OF SCIENTISTS IN LIFE SCIENCES (ICSLS)
Update 3rd December 2020 ~ Click- Cycle threshold (CT) values and their reference ranges, as applicable, must be reported by laboratories to Florida Department of Health (FDOH) via electronic laboratory reporting or by fax immediately.
Update 15th December 2020 ~ Click- Cease and desist papers served on Prof. Dr. Christian Drosten by Dr. Reiner Füllmich
Update 10th January 2021 ~ (Includes a section on PCR) Click - Request for Expedited Investigation Into Scientific Fraud in Covid-19 Public Health Policies
Update 20th January 2021 ~ WHO issues revised guidance Click ~ "careful interpretation of weak positive results is needed (1). The cycle threshold (Ct) needed to detect virus is inversely proportional to the patient’s viral load."
Update 1st May 2021. Dr John Lee, Prof of Pathology UK. Click ~ Unlocked Documentary
Update 15th September 2021. Reiner Fuellmich ~ Summary of findings of the Corona Investigative Committee
In conclusion -
SARS-cov2 is a multi-faceted problem which is currently affecting the world's population. Only one aspect has been considered here, namely, the cycle thresholds used in RT-PCR.
The relevance of cycle thresholds in the research technique known as Polymerase Chain Reaction (PCR) and in RT-PCR is unequivocal because it is an integral part of the process.
However, what seems to be lacking is an unambiguous range of what cycle threshold(s) should be utilised for the best outcome for the majority of people. Those who are at risk of becoming seriously ill require to be closely monitored and treated appropriately. Namely, quarantine and treat the sick because this should help prevent spreading to other vulnerable people. At the same time, those who are not at risk and pose very little or no risk to others should have no impediments to their lives and therefore contribute to the well-being of society, whether they are health care workers or teachers or any other profession.
It is hoped that those who have influence in setting public health strategies might want to consider and debate the issue of cycle thresholds as well as issue advice on preventative measures like Vitamin D3.
There is (and has been) a great deal of highly credible evidence from medical experts in this field that Vitamin D3 (at a certain level of intake) is a good preventative measure. For more information about the benefits of vitamin D3, including dosage, please click for this source ~ Vitamin D3 Evidence for immunity
See also, this most thought provoking piece of work ~ I had a Dream by David Grimes
There seems to be a fashion for tiers or levels at the moment, certainly with regard to social restrictions, lock-downs etc. Perhaps the public health strategists may wish to consider a 2-tier or even 3-tier strategy (traffic light scenario) when it comes to cycle thresholds?
For example, in conjunction with a medical diagnosis from a doctor, a positive RT-PCR up to 24 cycles and demonstrating physical signs and symptoms could be classed as (red light), quarantine and monitor/treat (depending on signs and symptoms) then re-test; from 25 to 34 cycles (amber) and demonstrating signs and symptoms, take sensible precautions e.g. off work for short period then re-test within short period of time; above 34 cycles (or very close to that = green light) no special action required except to act 'sensibly', re-test. Of course, I am not advocating this - It is simply substantial food for thought and worthy of consideration to instigate urgently needed research by those who actually do make the public health strategies, which in turn affect all of our lives.
It is one thing to talk about the nature of PCR, RT-PCR, and cycle thresholds, but it is another subject altogether to ask what exactly are the tests trying to look for?
The above list of sources is simply a short sample of what is available on the internet.
There appears to be an abundance of evidence to at least give the time to seriously debate the subject, particularly if you have influence or input to public health strategies.
When referencing any material on this page, please do NOT reference this page itself, but rather cite the material from the original source(s).
I hope this page will inspire YOU to do some research into cycle thresholds in RT-PCR. Finally...
|
The 'Great Barrington Declaration' extract...
"As infectious disease epidemiologists and public health scientists we have grave concerns about the damaging physical and mental health impacts of the prevailing COVID-19 policies, and recommend an approach we call Focused Protection."
However, no mention is made of cycle thresholds on this declaration (nor the efficacy of Vitamin D3). The declaration at this moment in time appears to be simply a mission statement awaiting the fleshing out of details. Perhaps the issue of cycle thresholds might be emphasised in the very near future, as well as Vitamin D3.
At the time of creating this page, the Great Barrington Declaration had received signatures from 30,821 medical practitioners, 11,051 medical & public health scientists, and 564,805 concerned citizens; these numbers are rising daily.
Please visit the website by clicking on the blue link below or by clicking on the photo. Alternatively, if you are cautious about clicking on links then you could type https://gbdeclaration.org into your browser. You have will have the opportunity to view videos, read the declaration, and if you wish, add your name to the growing list of concerned doctors, health professionals, academics, and members of the public...
Click here - Great Barrington Declaration
Thank you for taking the time to read this open letter.
|
*** End of page ***
|