Thank you for the heads-up on this Jack.

"It was approved by the FDA on the basis of clinical trial results
demonstrating test sensitivity of 100% and specificity of over 99.7%."

Did the FDA's clinical trial results calibrate the sensitivity/specificity
of the rapid test against the viral gold standard: that is, against an
isolate of the HIV virus itself? Alternatively: was the rapid test
calibrated against any test which itself has been calibrated against an
isolate of the HIV virus ?

To answer my own question: after twenty years there is still no test for
HIV that has been calibrated against the viral isolate.

Without reference to this "gold standard" we can know nothing about a
viral test's specificity. Without gold standard calibration the FDA's
figures are strictly meaningless. Without calibration every reactive HIV
test ever carried out might be a false positive. Calibration against viral
isolate is *absolutely required* .

Newcomers to this board may wonder why someone just doesn't calibrate a
test, any test, against an isolate of HIV.

There is a really good reason why no-one has ever done this. No-one has
yet managed to isolate the virus.

We have to ask ourselves why twenty years of extremely well-funded
research the AIDs industry has failed to produce a candidate virus
particle and then show that that particle is able to replicate in a virus
like manner. This is a basic first step of clinical epidemiology and is
conspicuous by its absence.

EVERYBODY REACTS POSITIVE
                                      ON THE ELISA TEST FOR HIV

                                                      By Roberto Giraldo

                                                   Continuum Midwinter
1998/9
 

       For the last 6 years I have been working at the laboratory of
clinical immunology in one of the most prestigious University
Hospitals in the
       City of New York. Here I have had the opportunity to personally
run and get to know in detail the current tests used for the
diagnosis of HIV
       status, namely, the ELISA, Western Blott and Viral Load tests.

       1. Diluting the serum for the ELISA test.

       The ELISA test is a test for antibodies against what is supposed
to be the Human Immunodeficiency Virus or HIV. To run
this test, an
       individual's serum has to be diluted to a ratio of 1:400 with a
special specimen diluent. According to the test kit manufacturer
this diluent
       contains

              0,1% triton X-100, Bovine and Goat Sera (minimum
concentration of 5%) and Human T-Lymphocyte Lysate (minimum
              titer 1:7500). Preservative: 0.1% Sodium Azide (1).

       This extraordinary high dilution of the person's serum [400 times]
took me by surprise. Most serologic tests that look for the
presence of
       antibodies against germs uses neat serum [undiluted]. For example,
the tests that look for antibodies to hepatitis A and B
viruses, rubella virus,
       syphillis, hystoplasma and cryptococus, to mention a few of them,
use straight serum [undiluted]. However, to try to prevent
false positive
       reactions some serologic tests use diluted serum; for example this
is the case with tests that look for antibodies to measles,
varicelia and mumps
       viruses which use a dilution of 1:16, to cytomegalovirus [CMV]
1:20 and to Epstein-Barr Virus [EBV] 1:10.

       The obvious questions are: What makes HIV so unique that the test
serum nedds to be diluted 400 times?. And what would
happen if the
       individual's serum is not diluted?.

       2. Testing the ELISA test without diluting the serum.

       To answer these questions I ran an experiment in a medical
laboratory in Yorktown Heights, New York. I ran it using the same
test kit reagents
       that are usually used to run the ELISA test in most clinical
laboratories worldwide (1).

       I first took samples of blood that, at 1:400 dilution, tested
negative for antibodies to HIV. I then ran the exact same serum
samples through the
       test again, but this time without diluting them. Tested straight,
they all came positive.

       Since that time I have run about 100 specimens and have always
gotten the same result. I even ran my own blood which, at
1:400 reacts
       negative. At 1:1 [undiluted] it reacted positive. I should mention
that with the exception of my own blood, the patient samples
all came from
       doctors who requested HIV tests. It is therefore likely that most
of the blood samples that I tested belonged to individuals at
risk for AIDS.

       According to Abbott Laboratories, the absorbance value [yellow
color intensity]

              develops in proportion to the amount of antibodies to HIV-1
which is bound to the bead (1).

       What I noticied is that the absorbance values of the specimens
that tested negative when diluted [1:400], but positive when
undiluted [1:1], had
       lower values than the samples that, diluted, react positive on
both the ELISA and Western Blott tests. This would probably
mean that the blood
       that is negative when diluted but positive when undiluted has a
lower level of antibodies than the diluted blood that is doubly
positive and,
       therefore, may probably test negative on the Western Blott test.
However, I have not had the opportunity to check this
hypothesis.

       The graphic below ilustrates how blood that reacts negative for
HIV at 1:400 ratio always turn positive when run at 1:1
[undiluted].
 
                              Run of ELISA test for HIV with two
different concentrations of the person's serum.
                                 (a) Results at 1:400
                                                                         
   (b) Results at 1:1
                              9112324b G5 0.076 ---
                                                                       
9112324b G5 0.262 reactive
                              9112325b H1 0.081 ---
                                                                       
9112325b H1 0.259 reactive
                              9112326b H2 0.071 ---
                                                                       
9112326b H2 0.329 reactive
                              9112327b H3 0.060 ---
                                                                       
9112327b H3 0.401 reactive
                              9112328b H4 0.073 ---
                                                                       
9112328b H4 0.345 reactive
                              9112329b H5 0.062 ---
                                                                       
9112329b H5 0.343 reactive
                              9112330b J1 0.060 ---
                                                                       
9112330b J1 0.234 reactive
                              9112331b J2 0.077 ---
                                                                       
9112331b J2 0.306 reactive
                              9112332b J3 0.067 ---
                                                                       
9112332b J3 0.248 reactive
                              9112333b J4 0.086 ---
                                                                       
9112333b J4 0.222 reactive
 

                                    Column (a) shows 10 specimens
reacting negative at 1:400 dilution.
                                   Column (b) shows the same specimens
reacting positive at 1:1 dilution.

       It is important to note that the Western Blott antibody test for
«HIV» also needs serum to diluted. Although it too has an
usually high dilution,
       here the individual serum is only diluted at the ratio of 1:50
(2). I have not yet had the opportunity to run this test with
undiluted [1:1]
       specimens.

       3. Discussion.

       The following are three possible explanations for why undiluted
specimens of blood always react positive at the ELISA test:

       3.1. Everybody has HIV antibodies.

       It is accepted worldwide that the ELISA test for HIV detects
antibodies against what is known as the Human
Immunodeficiency Virus
       (3-4-5-6). And the pharmaceutical company that commercialises the
ELISA kits states that

              Abbott HIVAB HIV-1 EIA is an vitro qualitative Enzyme
Immunoassay for the Detection of Antibody to Human
              Immunodeficiency Virus Type 1 (HIV-1) in Human Serum and
Plasma (1).

       Since all undiluted blood specimens react positive on the ELISA
test, a test that supposedly tests for antibodies to HIV, the
results presented
       here suggest that every single human being has HIV antibodies. And
this suggests that everybody has been exposed to HIV
antigens.

       This would mean that all of us have been exposed to the virus that
is believed to be the cause of AIDS. The people that react
positive even at a
       dilution of 1:400, would be the ones that have had the highest
level of exposure to HIV antigens. The rest of the people -the
ones that only react
       positive with undiluted serum [1:1]- would have had a lower level
of exposure to HIV.

       3.2. Everybody has different levels of HIV infection.

       It is also believed worldwide that a person that reacts positive
for antibodies against HIV has not only been exposed to but is
infected with a
       deadly virus that causes immunodeficiency (3-4-5-6). Therefore,
the positive reactions of all undiluted sera would mean that
everybody, or at
       least all the blood samples that I have tested, including my own,
infected with this «deadly» virus. The ones that react positive
at a ratio of 1:400
       would simply have a higher level of «deadly» infection than the
«deadly» infection has by the ones that reacts positive only
with undiluted
       serum.

       3.3. The test is not specific for HIV.

       The results presented here could also mean that the tests used for
detecting antibodies to HIV are not specific for HIV, as has
been explained
       previously (7-8-9-10-11-12-13-14). In this case, there would be
reasons other than HIV infection, past or present, to explain
why a person
       reacts positive to it. The test also reacts positive in the
absence of HIV (7-8-9-10-11-12-13-14).

       The scientific literature has documented more than 70 different
reasons for getting a positive reaction other than past or present
infection with
       HIV (7,10,11,14,15). All these conditions have in common a history
of polyantigenic stimulations (15,16).

       Even Abbott Laboratories is well aware of the specificity problems
with the ELISA tesst. This is why they state:

              EIA testing alone cannot be used to diagnose AIDS, even if
the recommended investigation of the reactive specimens
              suggests a high probability that the antibody to HIV-1 is
present

       and

              Although for all clinical and public health applications of
the EIA both the degree of risk for HIV-1 infection of the
person
              studied and the degree of reactivity of the serum may be of
value in interpreting the test, these correlations are impefect.
              Therefore, in most settings it is appropiate to investigate
repeatably reactive specimens by additional more specific or
              supplemental tests (1).

       Interestingly, there are countries like Great Britain where the
diagnosis of HIV status is based on the ELISA test alone. No
Western Blott or
       any other test is needed there.

       The only proper way for establishing the sensitivity and
specificity of a given test is with a gold standard. However, since HIV
has never been
       isolated as an independent purified viral entity (17-18-19), there
cannot be a gold standard for HIV. The sensitivity and
specificity of the
       antibody tests for HIV have instead been defined based on the
assumption that HIV is the cause of AIDS. In this way,

              The Abbot studies show that: Sensitivity based on an
assumed 100% prevalence of HIV-1 antibody in AIDS patients is
              estimated to be 100% (144 patients tested).

       and

              Specificity based on the assumed zero prevalence of HIV-1
in random donors is estimated to be 99.9/o (4.777 random
              donors tested (1)).

              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).
                                                                         
                      [Emphasis is mine].

       Since there is no scientific evidence that the ELISA test is
specific for HIV antibodies, a reactive ELISA test at any
concentration of the serum
       would mean presence of nonspecific or polyspecific antibodies
(20). These antibodies could be present in all blood samples.
They are most
       likely a result of the stress response, having no relation to any
retrovirus, let alone HIV (21,22). In this case, a reactive test
could be a measure
       of the degree of one's exposure to stressor or oxidizing agents
(15,16).

       The inevitable conclusion is that all positive reactions for
antibodies to HIV are simply false positives. If nobody is positive for
HIV, then
       people who react positive on the ELISA test do so due to something
other than HIV.

       4. Proposal to find out the real meaning of the «HIV antibody»
tests.

       To uncover the meaning of these tests I propose a simple
experiment: Take blood from three groups of a people and run the
tests highly
       diluted, undiluted and at a wide spectrum of dilutions in between.
The first group would be a group of healthy people of many
age groups; the
       second group would be a group of people from the conventional AIDS
«risk groups»; the third group would be a group of
people with clinical
       conditions both related and unrelated to AIDS. All groups would be
subjected to both the ELISA and Western Blott tests.

       Additionally, all blood samples could be subjected to «the viral
load test for HIV».

       The results of such an experiment could determine whether these
test measurements bear any relationship to an individual's
level of exposure to
       stressor or oxidizing agents. If so, the tests could be salvaged
as a measure of an individual's level of intoxication.

       Let us find the economic support necessary to run this experiment.
In the mean time, since people are reacting positive on tests
that are not
       specific for HIV, let's please stop labeling them as «HIV
positive».

       Acknowledgments.

       I want to thank Mr. Albert Padovani, Director of Yorktown Medical
Laboratory for permitting me to run the experiments
reported here in his laboratory and for
       providing the reagents for the tests. Also I thank Tom Di
Ferdinando Executive Director of Health Education AIDS Liaison
(HEAL) in New York City for editing the
       manuscript for this article and for his valuable suggestions.

       Roberto A. Giraldo, MD. Physician, specialist in internal
medicine, infectious and tropical diseases. Member of the Boards od
Directors of
       The Group for the Scientific Reappraisal of the HIV-AIDS
Hypothesis and Health Education AIDS Liaison (HEAL).
Independent AIDS
       Researcher. Author of the book AIDS and Stressors, New York City.
E-mail: rgiraldo@cdiusa.com.

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