Infectious Disease Postdoc/Clinician
Department of Pediatrics, University Hospital, Syracuse NY
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The Perth Group have rather missed the point.
If uninfected cells have a value of X and infected cells have a value of Y then surely it's obvious that Y-X is the contribution of viral RT activity...?
As to "Nobody has ever suggested Nicholas Bennett's "method". " I'm afraid I cannot lay claim to this, since it's standard laboratory practice .
The Perth Group are also apparently unaware of the use of isoenzymes (CK-MB versus total CK) to distinguish myocardial infarction from muscle destruction. To quote from an online source :
"If the ratio of CK-MB to total CK (relative index) is more than 2.5- 3, theheart is the likely muscle damaged. A high CK with a very low relative index suggests that other muscles were damaged."
"Repeat, scientifically it IS possible to determine what is the cause of reverse transcriptase activity (or raised CK) on the basis of "RELATIVE amount".
Other methods to distinguish retroviral RT from cellular enzymes include the use of actinomycin D, varying salts and pH's and column purification. To suggest otherwise is blatantly lying, since this has been explained to the Perth Group on this forum repeatedly. Such methods were employed by Montagnier in the "isolation" paper.
I'm curious as to the actual basis for their statement that "In Montagnier's 1984 paper the level of RT activity by the DNA polymerases beta and gamma is at least as high as that of the "HIV" polymerase." According to the methods I use, that clearly is not true. I'm more inclined to suggest to the reader that, based on their previous track record, the Perth Group are either misunderstanding or misrepresenting the true data. Could the Perth Group come up with an alternative explanation for consistently low RT values in uninfected cells versus identially treated infected/transfected cultures?
If the Perth Group persist in refusing to educate themselves, could they at least keep their misguided opinions to themselves? Surely it's the purpose of the BMJ Rapid Responses to allow valid criticism of recent publications, and not to be misused as a soapbox for promoting false information. It is quite clear from this post and the many others that have appeared here in the last year that the Perth Group have no intention of stopping their attempts to convince the lay reader of the innocuous nature of HIV. They are currently flailing so badly it isn't even an effort to correct them. May I suggest to the editors that a measure of moderation be employed, before someone dies from believing them?
As regards their ignorance of retroelements, it would be obvious to anyone who knew their genetic makeup that they include many of the features of exogenous retroviruses: tRNA primer sites, poly A tails, transcription/translation signals etc etc etc. Practically the only consistent lack is a packaging signal on the RNA or an envelope gene (but Syncitin of course is the prime example of a retroelement that contributes an envelope gene to normal human physiology). I did not exclude them, but rather specifically included them! Transfection of course is a whole different ball game, since it is usually is of DNA and not RNA, so the reverse-transcription and integration stages aren't applicable. The constructs usually include the appropriate transcription/translation signals (as created in the lab) and so the other criteria are also fulfilled. I had hoped they wouldn't fall into the rather obvious trap of displaying their ignorance in this way: and no doubt if I had said "Yes" in any other place they would have said "How therefore can you exclude...." They have a (pseudo)answer for everything, and yet nothing.
I note their recent attempt to rebutt my prasee of the mechanism by which HIV causes AIDS. I will be only too happy to provide appropriate references and explanations in due course.
1. Potts, B. J. 1990. ''Mini'' reverse transcriptase (RT) assay, p. 103-106. In A. Aldovini and B. D. Walker (ed.), Techniques in HIV research. Stockton Press, New York.
Competing interests: None declared