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Re: Re: Re: Out of all the Responses Four Arguments Have Emerged. Three of These Things are not Like the Other...

Because a Western Blot test is also an antibody test, how does Brian Foley know this 'confirmatory' test is not also registering as false positives?

Brian Foley wrote:
"There are many “gold standards” for antibody tests."

How can there be 'many' gold standards for an antibody test? Do all these gold standards agree among themselves? Would logic not dictate that the only valid gold standard be the determination of the actual presence or absence of the thing you are testing for, in this case virions of 'HIV'. That is, prove that people with a reaction on the antibody tests have virions, proven to be 'HIV,' in fresh, uncultured blood. It would also seem that any gold standard could only be isolation, otherwise it would, in and of itself, need a gold standard. For instance, detection of reverse transcription activity, PCR genetic technologies, P24 antigen, antibody/antigen reactions are all not detection of actual virus and are proned to misleading readings.

If there are so many gold standards, why are they not used for the HIV antibody tests? For instance, Abbot Laboratories says of their ELISA test, 'At present there is no recognized standard for establishing the presence and absence of HIV-1 antibody in human blood. Therefore sensitivity was computed based on the clinical diagnosis of AIDS and specificity based on random donors"1

Perhaps Brian Foley can give 'many' of these gold standards to Abbot Laboratories whereupon proper determination of the sensitivity and specificity of their tests can be made without having to resort to assumptions.

In addition, the Western Blot test, which Brian Foley says can confirm a pregnant woman's infection status, says "Do not use this kit as the sole basis of diagnosing HIV-1 infection"2 Does this mean that if a western blot test is fully reactive in a person, and this person does not receive an EIA test (which is considered the least specific 'HIV' test), then the WB is not valid?

Brian Foley wrote:
"There are dozens of different methods for distinguishing true infection from false positive results.

What are these dozens of different methods and are they used routinely on people with a positive western blot test? How do these dozens of different methods determine the actual presence or absence of 'HIV' so as to give proof of a 'true' positive?

Brian Foley wrote:
"It is for this very reason that countries which can afford to do so use a very sensitive test such as EIA for initial screening, followed by a more extensive test such as western blot on those samples which react positive to the initial EIA."

It's my understanding that the United Kingdom does not routinely use the Western Blot test to 'confirm' reactive ELISA tests. Does this then mean that reactions in England may or may not be considered 'real' infections in places that do use the Western Blot. The same for places like Africa. If a person is given a positive ELISA test result, but it is not 'confirmed' by WB, how can they be sure it's a 'real' positive?

Brian Foley wrote:
"Each country has its own medical councils and governing bodies, comparable to the USA’s CDC, NIH and FDA. The World Health Organization can make recommendations, but in the end each country is free to define its own standards for both diagnosis of infections (whether it be HIV or TB or Giardia) and for treatment of people who are diagnosed as having those infections."

If a person from a country whose governing body has determined that a single EIA test is sufficient to prove infection, is given a test and found positive, is this as valid as a 'fully confirmed' by western blot infection in a wealthy country? If so, why do wealthy countries bother with a western blot? Is this also sufficient proof to administer a pregnant African woman AZT or Navirapine? After all, the only thing concerning the average person receiving one of these tests is, do I truly have this virus? In 1987 former US Senator Lawton Chiles of Florida told an AIDS conference how twenty two blood donors were informed they were HIV infected on the basis of an ELISA test, whereupon seven then committed suicide. Were or were not these people infected by 'HIV'? Since nowadays one cannot receive a 'positive' diagnosis on the basis of a single ELISA/EIA, does this mean that a person coming from Africa having received a 'positive' diagnosis via this test must then be 'confirmed' by Western Blot in order to be a valid 'positive' in the US? If this person happens to have a negative Western Blot, that is, this person's 'HIV infection' is not 'confirmed', does this person get to stay 'HIV negative' upon returning to Africa?

Additionally, if a person in Africa is given a presumptive diagnosis of AIDS (in other words, no serology is performed), is this also proof of infection?

Brian Foley wrote:
"Again, if you look at the differences in criteria for diagnosis of HIV infection, and you look at actual EIA and western blot results from thousands of people, you will find that a very small percentage of actual test results fall into the range where they would be labeled as “indeterminate” in one country and “infected” in another. It would be foolish to assume that any test that is attempting to give a binary “yes or no” answer to a question that is based on an analog signal (After infection, most infected hosts begin to produce antibodies, and the level of antibody production rises over time. It does not instantaneously jump from zero antibody to a high level.) to NOT have a rather arbitrarily set point where values below are labeled as “no” and values above are labeled as “yes”." It is true we are essentially testing an analog signal, however, at issue is whether one of these 'weak' or 'strong' signals is synonymous with the actual presence of that which is being tested for. How many people after having been told they have an initial positive reaction (that is, they are 'weakly' reactive yet still 'positive') are then advised to go back and receive further testing until such time that their 'many' antibodies 'prove' true infection? Instead I believe a person is told they are 'infected' as long as their test meets an arbitrary criteria and then go on believing that from then on they are infected. For instance, the US Consortium for Retrovirus Serology Standardization reported that 127/1306 (10%) of individuals at "low risk" for AIDS including "specimens from blood donor centers" had a positive HIV antibody test by the "most stringent" US WB criteria. Were these at low risk individuals told they were infected or were they advised to continue testing until they no longer satisfied the infection criteria? If a person has a Western Blot test result of p24, p31/32 and gp41, should that person be treated immediately with drugs like AZT and various protease inhibitors (whose toxicity profiles are well established)? Or should they be told to wait until all bands have become positive such that their infection status is 'proven' by 'many' antibodies and not simply 'transient' reactions?

Brian Foley wrote:
"Most EIA and western blot results that would be labeled as “indeterminate” in the USA are transient" In the MACS, one band was considered proof of HIV infection in gay men until 1990. In men considered positive by one band, were tests repeatedly done to determine if they were 'transient'? Or were they considered true simply on the basis of being in a risk group?

Regardless, having many reactive bands on a western blot test does not prove that they were caused by 'HIV' antibodies. If one band can cross react, then all can be cross/non-specific reactions. If one considers that the groups said to be most at risk for AIDS are exposed to agents and situations that are known to cause high levels of circulating immune complexes, would it not follow that these groups are more likely to have higher levels of cross reactions? Again, without viral isolation as a gold standard, how can one tell those who are 'truly' infected from those who are not?

In a June 20th, 2003 post, 'Distinguishing between true and 'official' HIV infection", the Perth Group wrote, "the crux of Foley’s interpretation revolves around the time of appearance and the numbers of "weak" or "strong" WB bands. Apparently "two weakly positive western blot bands" may or may not be HIV but "a strong ELISA positive plus 3 or 4 strongly reactive western blot bands" definitely are HIV. Is this system ex cathedra or does Brian have proof? If he has proof what is it?"

And, in the same post, "As mentioned, Brian’s criteria involve arbitrary appearances and numbers of “weak” and “strong” ELISAs and WB bands. It goes without saying this is not proof of antibody test specificity. If Brian were handed a small object resembling an apple, does giving him a ton of the same but larger three weeks later prove it truly is an apple?

Brian Foley wrote:
"Infectious molecular clones of dozens of different isolates of HIV-1, HIV -2 and many different SIVs have been created and used in research. HIV-1 has thereby been shown to be a group or family of related viruses most closely related to the lentiviruses isolated from Chimpanzees."

and;

"Accurate classification of viruses cannot be done by electron microscopy, EIAV looks identical to HIV-1, and a mixture or EIAV and HIV-1 cannot be distinguished from pure HIV-1 by electron microscopy."

If 'HIV' is a lentivirus, why did Montagnier initially claim it was a Type C retrovirus based on EM's of cell cultures now said to have been infected with 'HIV', a lentivirus? In addition, 'infectious molecular clones' rely on cloning something. One cannot clone something from nothing. Cloning a clone creates a clone and says nothing of its origin. This something, at one point in time, had to be identified for the very first time.

In the early 80's several scientists believed they might have a retroviral entity in cell cultures which they also believed might be the 'probable cause of AIDS'. They extracted materials (proteins and RNA) from sucrose density gradients and claimed they belonged to a new, externally acquired molecular, retroviral entity, HIV (LAV/HTLV-III at the time). How did they prove that these materials belonged to a new retroviral particle, 'HIV'? If some proteins in this gradient reacted with antibodies from gay men with AIDS, how does this prove these proteins were constituents of this particle?

Perhaps a scientist cannot distinguish between several different forms of retroviruses based on EM's of particles. But, AT LEAST one should see retroviral particles fulfilling the morphology of the particles which are now claimed to have been isolated in this density gradient. For instance, even in the now defunct HL23V, Gallo published EM's of virus?like particles banding at a sucrose density of 1.16 gm/ml. Did Gallo and Montagnier publish EM's of the 1.16 gm/ml band showing lentivirus like particles to prove they AT LEAST had the possibility of a retrovirus in hand. If not, how did they prove these proteins and RNA were constituents of this new retrovirus? If they extracted RNA and claimed it was the genome of a new retrovirus, but there were no particles in this gradient, what then has the scientific community been cloning?

Brian Foley continues:
"Serological methods are much better, but the true "gold standard" is cloning and sequencing infectious molecular clones."

Forgive my ignorance here, but could one pass any stretch of DNA through a bacterial vector and call it 'infectious'?

It would again seem to me that serology, that is, antibody/antigen reactions, are only as good as their gold standard. One cannot reason backwards with an antibody reaction and so it must have a reference standard against which its reactions can be compared. And since a clone is only as good as its source; and since the sources of these clones have their origins in 1983/84 where neither research group showed in their gradients particles fulfilling the morphology of a lentivirus, I fail to see how it can be used as a gold standard for this serology.

Perhaps I'm wrong, but if 1000 individuals are said to 'infected with HIV' (in this case let's assume by having all bands reactive on a western blot), should it then not follow that retroviral particles fulfilling the morphology of a lentivirus can be seen under an electron microscope from fresh, uncultured blood taken from these individuals? Should it not be possible to extract entire genomes of these virions from this fresh, uncultured blood? In addition, if a person is given a PCR viral load test, and is told they have a very high viral load, can then fresh blood be taken from this person and examined by EM to see this multitude of active virions?

Brian Foley wrote:
The answer is that while PCR has a very low false-negative rate, it can have an unacceptably high false positive rate. In the case of newborns, the infant has maternal antibodies in its bloodstream which make it difficult to distinguish infected infants by antibody methods.

If PCR is sufficient to diagnose 'HIV' infection in infants 18 months of age or less, why would it not be sufficient to 'prove' infection in anybody of any age? Surely, if the PCR primers are so highly specific to 'HIV' then it should be no problem to use for diagnostic purposes in all people. Conversely, if the PCR test is not good enough to diagnose an adult, how can it be good enough to diagnose an infant? As with so many other 'HIV' tests, the FDA doesn't feel sufficiently confident to give them diagnostic approval, only prognostic status. The Amplicor HIV-1 Monitor test kit says, "The Amplicor HIV-1 Monitor test is not intended to be used as a screening test for HIV or as a diagnostic test to confirm the presence of HIV infection"3

It makes no sense that a test, whose claim is the ability to detect 'specific' genetic material to 'HIV', can be used to determine viral load, yet at the same time cannot be use to determine the actual presence of 'HIV'.

Finally, how can a test with 'an unacceptably high false positive rate' be used to accurately determine 'viral load'?

Chris Tyler

1. Abbott Laboratories, Diagnostic Division, 66-8805/R5; January, 1997 2. HIV-1 Western Blot Kit, Epitope, Inc., Organon Teknika Corporation PN201 -3039 Revision #8 3. Roche Diagnostic Systems, Inc., Amplicor HIV-1 Monitor Test Kit. US: 83088. June 1996)(13-06-83088-001

Competing interests: None declared