HIV EM and purification 28 June 2004
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Nicholas Bennett,
Infectious Disease Postdoc/Clinician
Department of Pediatrics, University Hospital, Syracuse NY 13084

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Re: HIV EM and purification

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 Characterization of highly purified, inactivated HIV-1 particles isolated by anion exchange chromatography.

The only caveat perhaps being that the virus particles where UV- inactivated prior to purification and analysis, which obviously resulted in RNA fragmentation. Other options include Optiprep centrifugation after sucrose centrifugation, which has been described extensively in the literature and used to demonstrate virally-incorporated cellular proteins.

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 1mm 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.

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 abscence, this point has been raised and addressed in public in other forums.

Calculations: 4/3*pi*r^3 for a 120nm diameter particle, into a 1mm cube gives a little over 1 trillion particles. Average blood volume taken to be 5 liters.

Competing interests: None declared