Department of Medical Physics, Royal Perth Hospital, Western Australia, 6001,
Valendar F Turner, John Papadimitriou, Barry Page, David Causer, Helman Alfonso, Sam Mhlongo, Todd Miller, Christian Fiala
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One AIDS patient will suffice
In his rapid response "Re: HIV EM and purification", 28th June 2004, Nicholas Bennett wrote: "Further to previous replies concerning published EMs of HIV, the best examples I have seen personally have been those in the paper: Richieri SP et al. Vaccine. 1998 Jan-Feb; 16 (2-3):119-29 Characterisation of highly purified, inactivated HIV-1 particles isolated by anion exchange chromatography".
In the photocopy of the Richieri et al EM that we have obtained it is not possible to distinguish any particles with the morphology attributed to "HIV". Would Nicholas Bennett please estimate what proportion of the material in the EM has morphological characteristics of retroviral particles. Would he please arrow some of the particles in the EM which in his view are HIV and email us a scanned copy.
Nicholas Bennett wrote: "In addition, an earlier post by the Perth Group asks why the supposedly ubiquitous virus cannot be isolated directly from the bloodstream of infected patients. Such a question displays a clear ignorance of the problems associated with virus isolation and purification at the most basic level. The size of the particle is such that 200 AIDS patients with viral loads of 1 million per ml would need to have their entire circulating blood volume concentrated to produce enough virus to fill a 1 mm cube (a grain of salt). Viruses are small, even if they can be prolific. The total RNA purified from that (bearing in mind that each virus contains 2 copies of the genome) is about 65 nanograms, or only just enough to visualise on a standard ethidium gel.
Asking for an EM from a virus that hasn’t been cultured or RNA from a virus that hasn't been amplified by PCR (or culture) is simply setting up a straw-man argument."
Nowhere in this debate or in any of our writings have we asked for proof of "HIV" purification and thus for the existence of the "HIV" genome (RNA) "directly from the bloodstream of infected patients". As far as evidence for the existence of "HIV" particles in the plasma is concerned, let us see what Hans Gelderblom, undoubtedly the best expert in "HIV" EM, has to say: "Routine two-step drop preparations for electron microscopic diagnostic procedures require particle concentrations of 106 to 108/mL. Therefore, negative evidence is not an absolute diagnosis. A number of effective concentration or immunologic procedures exist that markedly increase sensitivity of electron microscopic diagnostics for samples with lower particle concentrations. These procedures take from 0.5 to 16 hours and are labor and training intensive. Viral research or diagnostic facilities generally have access to at least one advanced procedure. Nonimmunologic procedures include: a) ultracentrifuge concentration—the material from cleared suspensions is sedimented by ultracentrifugation, resuspended in a smaller volume and then prepared by the standard two-step drop method; b) agar diffusion—a 20–50 [20- to 50-µL drop of suspension is placed on 1% agar. As the fluid is absorbed the virus is concentrated. After 15–20 min, a grid is placed on the remaining suspension and then stained as with the two-step method above (Figure 4D). This procedure will result in an enrichment factor of approximately 5x; and c) direct centrifugation to the electron microscopic grid with the Beckman Airfuge (Beckman, Palo Alto, CA) EM-90 rotor or A-100 rotor, a procedure that increases sensitivity up to 1,000 fold" .
Since: (i) there are AIDS patients who have particle concentrations of 106/ml ; (ii) "electron microscopic diagnostics for samples with lower particle concentrations" do exist; (iii) according to Robert Gallo more is known about "HIV" than any other virus; why to date has nobody published EM evidence for the existence of "HIV" particles in the plasma of even one AIDS patient?
Nicholas Bennett wrote: "I would also question their criticisms of Montagnier's RT data, which is in any case concentrating on a point that is over 2 decades old and frankly mis-interpreted anyway. In their conspicuous absence, this point has been raised and addressed in public in other forums".
Would Nicholas Bennett please: (i) tell us on what grounds he questions our "criticisms of Montagnier's RT data"; (ii) why is "that is over 2 decades old" relevant; (iii) why we have misinterpreted it; (iv) in what "other forums", "this point has been raised and addressed”.
1. Hazelton P, Gelderblom HR. Electron Microscopy for Rapid Diagnosis of Infectious Agents in Emergent Situations. 2003. www.cdc.gov/ncidod/eid/vol9no3/02-0327.htm
Competing interests: None declared