PCR FOR THE SO-CALLED MEASUREMENT OF HIV VIRAL LOADThe idea of looking for retroviruses in the blood of AIDS patients was in a way logical, because it is indeed in the blood of mice and chickens that zillions of retroviruses (RNA tumor viruses) were readily isolated and purified 40 years ago...
BUT:In all the old work (1960s) on experimental animals, the INITIAL step was the isolation of RETROVIRAL PARTICLES, either by sucrose density centrifugation, or (as in my work on the Friend virus) by two steps of Millipore ultrafiltration. This was leading, by a final, high speed centrifugation to a minuscule pellet that could readily be prepared for electron microscopy (transmission EM, plastic embedding, and thin sectioning), pellets in which thousands of packed retroviral particles could be easily demonstrated (see my 1997-78, vol 5 n°2, paper in Continuum, page 24), and pellets that could be then used for biological experiments (transmission of the disease to receptive experimental animals), and/or biochemical analysis (characterization of proteins and nucleic acids). The retroviral origin of these proteins and nucleic acids was unquestionable, because of the extremely high level of purity, demonstrated by EM, of the viral pellets. In all such experiments, all erythrocytes and leucocytes were first completely eliminated, by low speed centrifugation.
In today's so-called "viral load" studies this logical approach to retroviral isolation is completely ignored.
Simply because
NOTHING IS DONE to first isolate retroviral particles!
Instead: the PCR "Viral load" method starts of by first collecting LEUCOCYTES! Not viral particles!
Leucocytes are indeed collected, their nuclei extracted, their nuclear envelopes dissolved with detergent, and their CHROMATIN prepared for nucleic acid amplification by PCR !!! In any chromatin samples their is little surprise to find nucleic acid!
BUT: 6% or more of the human genome has striking homology with retroviral genome, a fact that is well documented for more than a decade. So, PCR has no difficulty to recognize short retroviral-like sequences in these human chromatin samples (never twice the same, but never mind: it keeps mutating !!), and to amplify it 1000 or million times! Bingo: this is HIV !!!!!! NO: it is the amplification of endogenous retroviral sequences that are present in ALL OF US! It has NOTHING to do with the hypothetical presence of circulating retroviral particles! It has nothing to do with any "measurement" of the "viral load". By that method, WE ALL have some level of ..."viral load"!!! Really? Yes, but to save the establishment from too much embarrassment NO CONTROL, on you and me, was ever made nor published! Do you know the reference of one single paper in which a large group of "normal" individuals would have been studied for HIV "viral load" by PCR measurement? I don't.
Add to this:
- That at Mbeki's conference, Pretoria May 2000, I formally stated that not one single retroviral particle has ever been visualized by electron microscopy in the blood of any patient with a so-called high viral load, and that that statement has never been refuted;
- That at the European Parliament debate, Brussels Dec 2003, I directly asked Luc Montagnier to give us his definition of the "viral load" and received an extremely ambiguous answer (page 196 of the proceedings), a fact that Prof. Gordon Stewart, who participated in that debate in Brussels, can most probably confirm.
In conclusion: the so-called measurements of HIV "viral load" by PCR methodology are completely missing any scientific relevance.
— Etienne de Harven, June 19, 2008