The Limits of Virus Isolation

By Julie Beal

The no-virus theorists have made people believe it’s possible to find a few viral particles in human body fluid (e.g. saliva or blood) and present them for all the world to see. When they say, ‘Show me your proof!’, they seem to be asking for a particle on a platter, as if it could be plucked from a sample with a pair of microscopic tweezers. Dr Cowan, for example, recently said, “It is not a technical problem to find a virus in bodily fluid”.

The no-virus theorists tell people it’s possible to purify body fluids so thoroughly, there should be nothing left but viruses. They say it’s been done with bacteriophages and giant viruses and it should therefore be possible to do it with saliva or blood, but they show no evidence to support this claim. At the same time, however, it makes perfect sense to expect that viruses should be taken directly from humans, and that they should always be purified. The fact that they’re not is definitely grounds for suspicion, so this article will attempt to explain some of the issues involved, from a purely physical point of view. It is part of a series of articles that provide a window into what virologists do, and why, but it is most certainly not an attempt to justify the use of vaccines. In fact, since germ theory has been put to bed, and since most viruses are harmless, there will be no expectation of disease, and no mention of Koch’s Postulates!

What’s the ‘right’ way to isolate viruses?

Stefan Lanka, who came up with the no-virus theory, says he managed to correctly isolate a giant virus[i] from an algae that grows in the ocean, and says it can also be done with phages (i.e. viruses that infect bacteria): “This is real, and cannot be contested. To isolate them, i.e. concentrate the particles, and separate them from all other components (=isolation), to photograph them immediately in the isolated state and to biochemically characterise them all in one go – this, however, has never happened with the alleged viruses of humans, animals and plants because these do not exist” (from ‘The Virus Misconception Part 1’). Lanka uses two definitions of isolation here. One is ‘separation from other components’, which is what purification is, and the other is ‘concentration of viral particles’, which can only be done after culturing viruses in cells of some kind. The marine virus that Lanka helped isolate in the mid-1990s was cultured in cells from algae, whilst phages[ii] are cultured in bacterial cells. Also known as ‘propagation’, the practice of culturing or growing viruses increases their number because they have a rather unique ability to produce clones of themselves. They keep on multiplying, or replicating, which means virologists end up with a concentrated sample of viruses. There needs to be a sufficient amount of viruses available so that after purifying them, there’s a usable sample remaining. Purification involves spinning cultured cells in a centrifuge to remove the cellular debris, but it usually ends up damaging a portion of the sample in question so there has to be enough material to start off with, so there’s enough left over to do tests on.

A study by Bachrach et al (‘Bacteriophage isolation from human saliva’, 2002) tested saliva for the presence of bacteriophages that are thought to “play an active and an important role in the oral environment”. Because they infect specific types of bacteria, phages can reduce the size of bacterial communities by killing them off, and this includes bacteria found in dental plaque and root canals. One of these is called Enterococcus faecalis, and 22% of the samples tested by the researchers contained the phages that infect this type of bacteria and which may therefore help limit their growth in the mouth. The researchers conclude by suggesting the phages could be used to prevent infections after root canal treatment.

To perform their study, “Stimulated saliva was collected from random donors who had not received antibiotics within the previous three months. One millilitre of each saliva sample was cleared of debris and bacteria by centrifugation at 15000 g for 10 min.” In order to isolate phages from the saliva, the researchers grew several strains of bacteria in Brain Heart Infusion broth (BHI), mixed them with the saliva, then vortexed the mixture briefly before incubating it for 20 minutes. Without purifying the mixture further, they prepared it for analysis by plating it on agar (a kind of sugar) together with more BHI (to feed it). The plates were incubated for five days or, “until lysis zones could be detected”. Lysis refers to the bacteria breaking down and dying due to the action of the phages, which is equivalent to the ‘cytopathic effect’ observed with most of the viruses studied by virologists. Lysis was seen in only one of the strains of bacteria they tested (i.e. Ent. Faecalis) which meant only phages that infect Ent. Faecalis were present in the saliva. (Antibiotics were not used in this study because they kill bacteria, but they are used when propagating viruses and exosomes, as described in previous articles and in the notes at the end of this one.)

Viruses are found in specific parts of the body

The only way to be sure a virus has come from a human is to study it in its natural habitat, and the ideal way to do this would be to obtain viruses directly from body fluids such as blood, urine and saliva. However, autopsies and other studies indicate that most viruses are tissue-specific, and only a few are blood-borne[iii], so samples are taken from the parts of the body that are usually affected, depending on the type of virus being hunted. SARS-CoV-2, for example, is said to commonly infect nasal cavities (and the lungs, etc.), and a study by Mondeja et al (2021)[iv] appears to show direct visual evidence of this[v]. The researchers describe using high-resolution microscopy to study the nasal cells of people who tested positive for the rona, and they did this, “directly in positive SARS-CoV-2 clinical samples rather than viruses grown in cell cultures”. They used nasal swabs from six people who tested negative for the rona, and had no symptoms, as well as swabs from six people who tested positive and also had ‘symptoms of covid’ (but not, it would seem, a significant disease). The cells were imaged using three different types of advanced microscopy techniques, and the ones who tested negative had normal healthy nasal cells whilst the ones who tested positive did not. The images of the apparently infected nasal cells showed there were, “characteristic coronavirus-like particles and evident destruction of microvilli”, which the researchers thought might explain why some people lost their sense of smell for a while. Dr Kaufman recently suggested[vi] that proof of a virus could only come from, “biopsies from individuals who are sick, showing tissue destruction”, which is what this study purports to show. Nonetheless, the evidence for the presence of the rona in these samples is relatively flimsy, because the researchers did not obtain full genome sequences and instead relied on electron microscopy, RT-PCR test results[vii] and very vague symptoms. Nor were the samples taken from actual biopsies! Sections of tissue removed from the lung, for example, provide a lot more material than nasal cells but are a lot harder to get hold of and also limited in scope because cells don’t survive for long after being removed from a living being.

In their book ‘Virus Mania’, Engelbrecht and Kohnlein say that HIV, Hepatitis C, H5N1 (bird flu) and other allegedly pathogenic viruses have not been proven to exist, “even though it’s as easy as taking a sample of patient blood and isolating one of these viruses, in a purified form with its complete genetic material (genome) and virus shell, directly from it, and then imaging it with an electron microscope.” There doesn’t seem to be any evidence to support this claim, and according to Karey Mullis, “[Dr Fauci] thinks you can take a blood sample and stick it in an electron microscope and if it’s got a virus in there you’ll know it. He doesn’t understand electron microscopy and he doesn’t understand medicine and he should not be in a position like he’s in.”

“The main problem with the electron microscope is that specimens must be fixed in plastic and viewed in a vacuum, and must therefore be dead. Other problems are that the specimens can be damaged by the electron beam and they must be stained with an electron-dense chemical (usually heavy metals like osmium, lead or gold).” There are plenty of electron microscope images of the rona available, but they’ve all been taken after propagating the virus in Vero cells, when there are enough of them to see (and compare to pictures of negative controls, i.e. uninfected Vero cells).[viii] There are also images that have been generated using atomic force microscopy, which can be used to examine viruses in 3D. Some viruses, apparently, can be detected directly in biological samples using transmission electron microscopy (TEM).  Herpesvirus and parapoxvirus have been imaged in the clear fluid found in skin lesions, rotavirus and adenovirus in faeces, and polyomavirus in a urine sample and in brain tissue.

The trouble with using saliva

Considering people are being tested for the rona by having a swab stuck down their throat, you’d think it’d be possible to get them to spit in a cup and then look for viruses with an electron microscope. Dr Kaufman and Dr Cowan often describe an encounter they had with a virologist who told them it would not be possible to ‘find a virus’ even if there was a great big load of saliva pooled together from 10,000 people. They suggest this anecdote proves their theory is correct, but maybe it’s because it would mean that, instead of just searching for a needle in a haystack, virologists would have to search for one or more needles in 10,000 haystacks. If only one sample is tested, and a full viral genome is found, there’s a high chance the virus is present, but if multiple samples are combined, each of the sequences that make up the genome could have come from a different person, so there would be no way of knowing if a whole virus had been found in any individual, and no way to perform other types of test.

The no-virus theorists say genetic sequencing is a fraud, and, by likening it to the RT-PCR test, they’ve made people think virologists just ‘invent’ genomes on a computer using very short sequences that can be made into anything they fancy.[ix]  Like the rest of their theory, this claim now seems more believable just because so many people are saying it,[x] but no evidence is presented to support it. This is worrying, because sequencing is the best evidence there is. It’s certainly way better than a picture from an electron microscope! The no-virus theorists have thus seriously undermined the ability of activists to deal with the onslaught of genetic interventions that are headed our way. Almost nothing will escape these interventions, because the industry is HUGE. The mainstream media didn’t warn us of what was coming, or what waits in store, but geneticists are ready to twist and pervert every aspect of the natural world, and saying they don’t know how to sequence a genome robs people of their power to avert this from happening.

What about exosomes then?

Dr Kaufman claims it’s possible to isolate exosomes from saliva, and says it should, therefore, be possible to isolate viruses from saliva, too, but it depends on what you mean by isolation! They are certainly not removed from body fluids and presented as pictures in the way Kaufman suggests they do. Exosomes are very similar to viruses, and the way they’re isolated from human beings is very similar to the way viruses are isolated, except that they’re unable to replicate in a cell culture. Exosomes can, however, be extracted from both patient samples and cell cultures using a variety of methods, e.g. magnetic beads, and biomarkers such as tetraspanins. They are then analysed using many of the same methods used by virologists, most of which are based on genetics and protein expression, e.g. microRNA analysis and proteomics.

“Currently, having a standard method for exosome isolation retaining its biological properties with increased yield and purity is a major challenge. The most commonly used method is differential ultracentrifugation but it has its own disadvantages, which include high time consumption, low yield due to disruption of exosome integrity, and high protein contaminants.”

Although it’s relatively easy to gather a sizeable quantity of exosomes from a cell culture, there are far fewer exosomes to be found in a sample of saliva. Over the last few years, scientists have figured out that it’s possible to isolate exosomes directly from saliva, but it’s not that easy. In one study, researchers got people to drool into a cup, then they purified/concentrated it using a centrifuge. This resulted in a small pellet which was analysed using various genetic techniques, such as RNA extraction, just like virologists do with viruses. Apparently, however, the “viscosity and cellular contamination of whole saliva make it a less than ideal medium for exosomal isolation”. What’s more, “Although whole saliva is relatively easy to obtain, it has significant disadvantages as a medium for the isolation of mRNA. Whole saliva contains hundreds of thousands of cells of different origin, as well as contaminants such as commensal bacteria….”. Another problem for researchers is that saliva is full of nucleases; these are enzymes that quickly destroy the RNA that makes up most exosomes and viruses.

Anyhow, if you centrifuge a load of drool, you’ll end up with a pellet that’s got exosomes in it because they purify at roughly the same band as viruses. There may or may not be some viral particles in there but how would anyone know? If they were looking for a previously catalogued viral genome, they could do genetic sequencing, like they do with swabs, but if they’re trying to find out whether it’s a virus or not (like when the rona was first discovered), they have to get it to replicate. This confirms it’s a virus and creates a bigger sample that can be studied. It also confirms they’re not exosomes, because exosomes do not replicate. Genetic sequencing provides fine-grained details about their physical composition and confirms that replication has taken place by proving they’re all very close copies of the original. (The first time virologists get a virus to grow/replicate in a cell culture, they call it ‘isolation’; after that it’s called ‘passaging’.)

In short, there’s more than one way to ‘find a virus’ but the main proof comes from replication and genetic sequencing, and when it comes to exosomes from patient samples, evidence comes from genetic techniques such as RNA extractions and flow cytometry, as well as images from an electron microscope.[xi]

When is purification necessary?

All of the things described above can only be achieved by using cells of some description, but cell cultures are known to contain various contaminants that make the sample impure, and they can only be removed by purification. However, it would appear that virologists and manufacturers are only expected to purify samples and isolates when:

• A new virus is discovered (e.g. HIV, SARS-CoV, SARS-CoV-2)
• Preparing a seed stock of viruses[xii] (for vaccines or research)
• Patenting a virus (there are at least three patents for SARS-CoV)

The rest of the time, virologists are content to confirm the identity of a virus by getting it to replicate and obtaining a fully sequenced genome. This can only be done if there are multiple long reads present in the sample being tested. Genetic sequencing does not work like the RT-PCR test and genomes are not ‘invented’ by stitching tiny sequences together. It’s based on detecting long genetic sequences, and if the sequences being looked for are not present, it’s taken to mean the virus is not present either.

As described in previous articles, the Chinese scientists who announced they’d found SARS-CoV-2 were the ones who followed purification protocols and performed all possible tests. After this initial ‘proof’, experiments carried out by other virologists are deemed to be corroborating the initial findings, and each other’s findings.

This might explain the responses received by Engelbrecht and Demeter[xiii] when they wrote to researchers who’d reported isolating and imaging the rona to ask if their electron micrographs showed, “the purified virus”, but apparently none of the researchers said “Yes”. The writers reported four one-line quotes[xiv] from the replies they received, and people often cite these replies as evidence the rona doesn’t exist. Here’s an example: “We could not estimate the degree of purification because we do not purify and concentrate the virus cultured in cells.” This quote relates to a very basic study by Han et al, where the only objective was to find out if the rona could be isolated from “putative patients” in Korea. They were simply confirming the presence of a virus that had already been evidenced, by saying it replicated, it had the same appearance, and its genome was confirmed to be more than 99.9% the same as the ones that had been reported since the initial announcement the previous month. It wasn’t meant to be a landmark study that demonstrated the existence of a brand new virus, because that had already been done by the researchers in China who ‘discovered’ it. The accepted protocol for new virus discovery, it would seem, only requires rigorous evidence to be provided by the discoverers. This does NOT include proving pathogencity, because this will be based mainly on evidence from epidemiology (ahem! ‘deaths with covid’, etc.!). The most a virologist can do is offer insights into the molecular nature of a virus and how it might interact with the body, based on lab tests and animal experiments. So, as with SARS-CoV in 2003, a few small groups of researchers were deemed to have established the existence of the rona and the genome they all agreed upon became the ‘reference genome’ to which all subsequent genomes were compared. The way virologists see it, the Chinese researchers isolated the virus in the standard way, and they used purification techniques throughout the process. Patients were screened for a number of viruses and bacteria. Antibodies were tested extensively. Mock-infected cell cultures were used as a control. Images were generated from an electron microscope. A range of genetic sequencing techniques were employed, and the genomes sequenced by three different labs were compared and found to be the same. It was all very neatly done and virologists couldn’t fault it, but most of them were very surprised by the genome of the virus itself. What happened during this period requires a lot more scrutiny, because it laid the very foundation upon which the whole scam depends, and appears to have been carefully stage-managed from the start. It was vital to establish a Patient Zero which would be used to model infectivity/transmission rates, and thus amplify the level of alarm, and it was just as important to time the release of genomic sequences, because this would affect the time taken to finalise the vax designs. All of them had been prepared in advance, and simply needed a few tiny tweaks to be ready for production.

But of course the anti-covidian community is a million miles away from all this… Nobody seems to notice, nobody seems to care, because ‘genetic sequencing is a fraud and there’s no virus anywhere’.

Notes:

[i]   This virus is known as Ectocarpus siliculosus virus-1 (EsV-1). It’s a phaeovirus with icosahedral shapes and it integrates one copy of its DNA into the genome of this specific type of algae. Lanka describes it as “a harmless virus”, but it is now known that environmental triggers can affect its reproductive capacity. “The viral genome remains latent in vegetative cells and is expressed in cells of the reproductive algal organs, sporangia and gametangia only when stimulated by, for example, changes in light composition and temperature.”

[ii]  Lanka believes that bacteria can be transformed into phages by undergoing metamorphosis.

[iii]   When it comes to blood-borne viruses, metagenomic analyses have found that healthy humans have a range of viruses in their blood. These are collectively referred to as the ‘blood virome’, and the most commonly found ones are Herpes viruses and Anello viruses. Apparently, we also contain about 31 billion bacteriophages which, “traffic human tissue and blood on a daily basis”. More information about these phages, and all the other stuff found in humans, might help us work out how viruses came to exist in the first place (perhaps they are a cellular response to illness).

[iv]  ‘SARS-CoV-2: preliminary study of infected human nasopharyngeal tissue by high resolution microscopy’ by Mondeja et al (2021)

[v]  The no-virus theorists repeatedly insist on receiving visual proof of a virus, but at the same time, they mock the images produced by virologists and often say they are mere ‘artefacts’ produced by the microscope or just made up by a graphic artist, and therefore not at all real. Sometimes they say they might be exosomes, although Dr Kaufman says, “there’s no way to know what the particles are”, and if that’s the case, how can they ever be ‘biochemically characterized’?

[vi]   In this video [at 13:47] ‘Is virology wrong? Dr Kaufman vs Dr Wodarg debate on Fuellmich’s corona committee & Dr Cowan comment’

[vii]  The RT-PCR tests were not performed by the researchers and it’s therefore unclear if the tests were performed correctly, e.g. how many cycles were used.

[viii]  Control experiments are often performed by virologists, as described in a previous article.

[ix] The no-virus theorists follow the ideas of Stefan Lanka, who appears to misunderstand the process of alignment, and perhaps confuses it with the RT-PCR test. He claims, incorrectly, that virologists, “always use very short pieces of nucleic acids, whose sequence consists of four molecules to determine them and call them sequences. From a multitude of millions of such specific, very short sequences, virologists mentally assemble a fictitious long genome strand with the help of complex computational and statistical methods. This process is called alignment.” (Lanka, ‘The causes of the corona crisis are clearly identified; Virologists who claim disease-causing viruses are science fraudsters and must be prosecuted’; Wissenschafft Plus magazine, April, 2020). What alignment actually involves is comparing lengthy sequences found in a sample to previously identified sequences, e.g. by checking the BLAST database. As shown in an example at Wikipedia, different species of mammals, all of whom make a histone protein, have very similar sequences that make this protein, and they only differ slightly. They can be aligned one under the other in rows, and the differences are highlighted as a column. (The example shows sequences from human, chimp, mouse, rat and cow.) This is also how virologists compare sequences of different coronaviruses, and how they define what type of virus it is they’ve discovered, e.g. SARS and the rona are almost the same. Genetic sequences are far too complex and numerous to be able to “mentally assemble” them into a genome, which is why computers are used, and the only way a computer knows how to do it is because they have been pre-programmed with a specific type of algorithm. The evidence builds when different researchers get the same results, and furthermore, when the sequences are artificially synthesized (like doing reverse engineering), they produce a ‘functional’ virus that has distinct biological properties that are comparable to the viruses produced in cell cultures.

[x]  It is also popular to quote a snippet from the CDC as proof the rona doesn’t exist, i.e. the bit that says, “Since no quantified virus isolates of the 2019-nCoV were available…”. The key word here is “quantified” and it means that, when preparing the RT-PCR test, the CDC did not have a stock of virus whose concentration was known (this is called the ‘titer’ and it’s a bit like saying how ‘strong’ or infectious the virus stock is). What the quote does show, however, is just how messed up everything is becoming (e.g. using artificially produced RNA). There’s nothing natural about any of it and the more unnatural it gets, the easier it is to ‘fix the results’ for alleged outbreaks and DIY-vaxines. Anyhow, the CDC produced an isolate in January, 2020, as described in a previous article, and many more have followed.

[xi]  Exosomes are continually released from cells in the body, and can be imaged with an electron microscope in saliva, breast milk and plasma, as shown in a study by Lässer et al (‘Human saliva, plasma and breast milk exosomes contain RNA: uptake by macrophages’, 2011). After purification, a small pellet of exosomes was removed from the centrifuge and then prepared for examination with an electron microscope: “The exosome pellet was resuspended in PBS and loaded onto formvar carbon coated grids … fixed in 2% paraformaldehyde and washed. The exosomes were immunostained with anti-CD63 antibody … or isotype control …followed by staining with a 10 nm gold-labelled secondary antibody (Sigma-Aldrich). The exosomes were subsequently fixed in 2.5% glutaraldehyde, washed, contrasted in 2% uranyl acetate and embedded in a mixture of uranyl acetate (0.8%) and methyl cellulose (0.13%). The preparations were examined in a LEO 912AB Omega electron microscope (Carl Zeiss NTS, Jena, Germany).” Would this be considered an acceptable technique by the no-virus theorists?

[xii]“  The FDA’s Guidance for Industry defines a viral seed as, “a live viral preparation of uniform composition … derived from a single culture process…”. This document is full of information (important clues at least!) about contamination and purification requirements for cell cultures and viral seeds used for vax production, including information about Vero cells and retroviruses. It suggests viral vaccine seeds can be purified, “by molecular cloning, serial passage in medium containing neutralizing antibody directed against the adventitious agent, or plaque purification”. In 21 CFR 600.3(r), purity is defined as the “relative freedom from extraneous matter in the finished product, whether or not harmful to the recipient or deleterious to the product.” Is purity always relative?

[xiii]  ‘COVID-19 PCR Tests Are Scientifically Meaningless; Though the whole world relies on RT-PCR to “diagnose” Sars-Cov-2 infection, the science is clear: they are not fit for purpose’ by Torsten Engelbrecht and Konstantin Demeter, June 2020

[xiv]  One of these studies was reported by the very first team in China, and it was led by George Gao, who attended Event 201! Engelbrecht and Demeter offer a small quote from the study author: “[We show] an image of sedimented virus particles, not purified ones.” This is taken to mean there was no attempt at purification, but the paper does say patient samples were, “centrifuged to remove cellular debris”, and when viruses started to grow, they were ultracentrifuged in order to sediment the virus particles, which is in fact a type of purification. It provides information on the ‘sedimentation coefficient’ (Svedberg value, S value).

Read much more about the science behind the coronavirus injections at Julie Beal’s archive.

Image: Pixabay

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