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Are we all POSITIVE ? Questions about the ELISA TEST for HIV

The “HIV causes Aids” Hypotheses is based upon a few test methods. One one these testing methods is the ELISA TEST. read what Dr. Roberto Giraldo has foud out...

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].   more...

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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