Antibody Tests (ELISA, Western Blot)

“sometimes people will have an antibody that will cross-react with some normal protein but the pattern of proteins that are unique, gp120, gp141, g44[?], [p]17, p6[?], these proteins are nominal to HIV…when you get antibodies from a person that react and give a pattern, almost always you will isolate HIV. Almost always that person is going to get AIDS and those proteins that are proven, as we did in our papers, with specific antibodies…we know are encoded by HIV and are certainly not normal cells…

[Lawyer] Turner goes on 'Nonetheless, HIV experts apparently believe there are proteins belonging to a retrovirus HIV and claim to use them to detect "HIV antibodies" and thus prove HIV infection. Even if there was proof these proteins are those of a purified infectious particle proven to be a retrovirus, the fact that patients have antibodies that react with these proteins is not proof the antibodies are caused by infection with HIV. This is because antibodies induced by a particular antigen react not only with that antigen but they also react with other antigens. This is a critically significant issue'. Could you comment on that?

[Gallo]Yes…that's why we added the western blot to the ELISA. The ELISA is very sensitive, it gives too much false positives…we in our papers told the scientific world screen with the ELISA but confirm with the western blot…there would be too many false positives with ELISA alone. Very sensitive. So yes, you get some cellular debris and you make it from antibodies reacting and you think that person is positive when the person won't be positive. Having said that, nonetheless, ELISA alone isn't bad…it just gives too many false positives…With the retrovirus by the way, you know 'antibody' means infection because once the retrovirus infects…it integrates antigens. Infection is for ever. You don't get an antibody and then it goes away. You get an antibody when you become infected and that antibody stays with you because you will always have the virus genes making some viral proteins. Almost always that is the case…Is it true around March 1984 Francis and Jim Curran of the CDC arranged a blinded test of HIV antibody test using tests from the Institute Pasteur, your laboratory and their own test.

[Lawyer] Is it true that a number of samples that the Institute Pasteur recorded as positive were recorded as indeterminate on your test?

[Gallo] I honestly don't remember details but something like that is probably true. I thought that the - see they weren't doing the Western blot there and they hired somebody out of my lab to help them, which didn't create friendships very much. Basically in any event they were doing ELISA and the Western blot and they were making it - we thought that the cut-off for the ELISA - the bar was too down so there would be too many false positives, so we had agreement on some that were not verifiable by Western blot, that is true.

[Lawyer] My instruction is that after you got their positive tests back your institution changed the results which you had previously had as indeterminate to positive.

[Gallo] I don't - I don't recall that being the case but if we did we made the sensitivity better and returned to the Western blots that's perfectly plausible.”

“EIA [Enzyme Immune Assay or ELISA] is commonly used as a screening assay for many infectious diseases, including HIV. These assays are used because they are highly sensitive and generally amenable to automation, facilitating high-volume testing…The small disadvantage of such a highly sensitive test is that the test produces false positives, the number and type of which vary with the assay used and the HIV prevalence in the tested population. All HIV diagnostic laboratories must confirm repeated EIA screen-positive results by a confirmatory assay, usually with Western blot. Laboratories may choose to first test with a second EIA assay, which uses a different part of the viral antigen for antibody capture, as part of their testing algorithm. Specimens that screen positive in the first assay but negative in the second assay should still be considered for confirmatory testing if the patient is symptomatic or high risk.”

“AIDS and AIDS-related conditions are clinical syndromes and their diagnosis can only be established clinically. EIA testing cannot be used to diagnose AIDS, even if the recommended investigation of reactive specimens suggests that the antibodies to HIV are present.”

“Western Blot assay should not be used as a screening test. WB should be viewed as a supplemental test which can be used to confirm positive results obtained from EIA. However: Specificity is less than that of EIA. A significant number of indeterminate blots are seen in low risk populations.”

“Even before the AIDS virus became the officially accepted cause of AIDS, the CDC had already made antibodies against the virus the only definitive criterion to diagnose any of the heterogeneous diseases as AIDS in 1985. Their unorthodox decision to use antibodies against the virus (normally functioning as a vaccine), instead of the virus, for the diagnosis of AIDS was based on the flawed analogy with some bacterial pathogens. For example, syphilis bacteria can be pathogenic despite the presence of antibodies, e.g. the Wassermann test for syphilis. But viruses are typically unable to enter cells in the presence of anti-viral antibodies the basis for the effectiveness of Jennerian vaccines. Because of the CDC’s decision, AIDS is diagnosed worldwide if antibody against (!) HIV, rather than HIV, is detectable in a patient along with any of the CDC’s 26 diseases. Since 1992 even low T-cell counts are diagnosed as a condition, termed “HIV/AIDS”, which is treatable with anti-HIV drugs provided it occurs in the presence of antibodies against HIV.”

“nonspecific reactions may occasionally be seen in specimens from people who have prior pregnancy, blood transfusion, or exposure to human cells or media containing cultured HIV antigen. Because of these and other potential nonspecific reactions, specimens reactive with the Vironostika HIV-1 Plus O Microelisa System assay should be confirmed with a confirmatory test ,e.g., Western Blot testing…antibody. In individuals at increased risk of infection, such as homosexual men, hemophiliacs, or intravenous drug users, repeatedly reactive specimens are usually found to contain antibodies to HIV by additional, more specific tests. However, when the ELISA is used to screen populations with a low prevalence of HIV infections, nonspecific reactions may be more common than specific reaction…Specimens found repeatedly reactive by ELISA and positive by additional, more specific tests are considered positive for antibodies to HIV-1. Clinical correlation is indicated with appropriate counseling, medical evaluation and possibly additional testing to decide whether a diagnosis of HIV infection is accurate.”

“If the French intended to use the CDC to prove they had found the cause of AIDS, Gallo would do the same…[after blinded testing of a number of samples] only 48 percent of Gallo's AIDS patients' samples were positive, compared to the Pasteur [Institute]'s 72 percent…Almost all Sarngadharan's [a member of Gallo’s lab] equivocal blood-testing scores, recorded as ‘plus/minus’, had come from AIDS patients'…Gallo thought the CDC should allow him to change his borderline results to positive. Pasteur hadn’t asked to change any of its results after the fact and Don Francis [of the CDC] was against allowing Gallo that extra advantage. [However] Jim Curran agreed to Gallo’s request…now Gallo and the French each scored 92 percent of the pre-AIDS patients positive. Among AIDS patients, the French had gotten 80 percent right to Gallo’s 78.”

“The third-generation EIA HIV-1 antibody test had a sensitivity of 77% [false negative rate of 23%] and a false-positive rate of 3%. All of the patients with a false-negative third-generation EIA antibody test had a positive p24 antigen test.”

“Antibodies directed to the env-encoded surface glyco-protein gp160 [of HIV] were detected in the cervicovaginal secretions of a small proportion of HIV-seronegative sex workers in Abidjan. In 2.9 to 12.3% of these women, depending on the test used, the anti-HIV antibodies were present in vaginal fluids that were free of contaminating semen. Since there is no established gold standard test, it is unclear which of these two proportions is the best estimate of the real prevalence rate of cervicovaginal anti-HIV antibodies in the absence of contaminating semen in HIV seronegative sex workers...The 25 HIV-1-seronegative sex workers with anti- HIV antibodies in their semen-free cervicovaginal secretions by both in-house ELISA and Seradyn Sentinel HIV-1 Urine EIA [ELISA] had no evidence of HIV-1 RNA in plasma. It is therefore unlikely that these antibodies are part of a primary HIV infection, although these women were not followed up...In the present study, increased sexual exposure was not associated with the presence of HIV-antibodies in cervicovaginal secretions, as measured by either of the two tests.”

“False-positive ELISA [antibody] test results can be caused by alloantibodies resulting from transfusions, transplantation, or pregnancy, autoimmune disorders, malignancies, alcoholic liver disease, or for reasons that are unclear…The WB [Western Blot antibody test] is not used as a screening tool because…it yields an unacceptably high percentage of indeterminate results.”

“The serological diagnosis of HIV infection is usually made on the basis of the detection of circulating antibodies specific for viral antigens gp41, gp120 and gp160. Despite using recombinant immunogenic oligopeptides, which improved the sensitivity and specificity of immunological tests, a number of both false-positive and false-negative reactions have been reported. Although the emergence of new viral serotypes or recent infection could be responsible, at least partly, for the low sensitivity of serological assays in detecting early antibody responses, false-positive results could be explained by crossreactions with unrelated antigens. Spehar and Strand recently demonstrated the cross- reactivity of anti-gp41 murine monoclonal antibodies with the human cytoskeletal protein alpha-actinin, and antibodies reacting with both the immunodominant region of HIV gp41 and alpha-actinin have been found in the sera of HIV-infected individuals...Only three out of 108 sera with IgG anti-Tricomonas vaginalis, and one out of 32 with IgM, tested positive for HIV using both kits.”

“The cutoff value [of this ELISA test] indicating a positive result is 0.500. Optical densities of 0.300 to 0.499 are indeterminate [often called ‘borderline’ or ‘weakly reactive’ and need to be retested. Values below 0.300 are considered to be negative. In most cases, a patient will be retested if the serum gives a positive result. If the ELISA retests are positive, the patient will then be retested by western blotting analysis.”]

“Samples giving discordant results [one positive, one negative] on the two ELISAs were retested by both test assays. If the results were still discordant, WB [Western Blot] analysis was done [making the WB the 'gold standard' for HIV seropositivity. No consideration is given to false positive results on the WB]…Wellcozyme HIV-1 recombinant ELISA showed 31 and 7 false positive reactions on initial and repeat testing respectively…After repeat testing on both assays eight repeatedly discordant samples were run on WB assay; four showed a single p24 band, while the rest gave no band on the WB. One sample with p17, p55 and gp 160 bands on the WB was considered indeterminate…None of the 120 randomly selected sera which were negative on both ELISAs showed a positive WB pattern [this information is used to claim test specificity of over 99% because eventually at least two tests that agree were found]”

“[Table A of this test kit label shows that 168 out of 364 people (75%) classified as low risk had at least one of 8 bands positive, but only 126 people were eventually classified as positive. Out of the 75% with at least one positive band only 54% had a p17 band while 79% had gp120/160. Amongst people with AIDS while 100% had a gp41 band and 98% had a gp120/160 band, only 21% had p17 and only 42% had p55]”

“Env-specific T helper responses evaluated in a total of 28 HIV seronegative, PCR negative HCW [health care workers] with HIV exposures [of which 10 had taken AZT], indicated that 21 of them (75%) showed responses to two or more of the five peptides [proteins believed to be from HIV] and in certain individuals these responses were observed more than 23 wk after exposure. Surprisingly, 24% (9/38) of HCW with HIV negative exposures exhibited responses to the peptides in contrast to 9% (3/33) in healthy blood donors…and the difference between the groups were statistically significant. It cannot be ruled out that some of the exposures to negative fluids were actually to fluids from HIV-infected individuals who had not yet seroconverted [or that the 'HIV' proteins are not actually specific for this virus]”

“The exact duration of the period between first contact with HIV and the appearance of specific antibodies varies and remains a subject of controversy. Seroconversion usually takes place 2-3 months after HIV infection…We report a case of HIV infection following a needlestick injury in which seroconversion was observed beyond the 6th month after the accident. A 47-year old woman…was accidentally pricked by a needle on May 10, 1993 at the clinic where she worked as a cleaner. Serological tests in accordance with French law were done at the time of the accident and in the 3rd and 6th month afterwards…She had no personal risk factors for HIV infection, and no at-risk circumstances between the accident and the diagnosis of HIV infection. Symptoms of possible acute primary infection were observed at the…6th month after the accident…HIV serology…was [first] positive at the 8th month. The first positive western blot showed a full pattern of infection. Serum p24 antigen remained negative on all studied samples.”

“Two women had a history of HIV seropositivity but were later found not to be infected.”

“The sensitivity and specificity of the human immunodeficiency virus (HIV) type 1 enzyme immunoassay (EIA) are greater than 99%... [but] between 4% and 20% of serum samples that are repeatedly reactive by HIV-1 EIA are interpreted as indeterminate by Western blot [which is used to confirm the EIA]...Of the 131 EIA-negative controls 35(27%) had indeterminate Western blots..Of 244 cases referred to the study because of previous reactive EIA and indeterminate Western blot, 139 still had repeatedly reactive visit 1, and of these, 124 also had indeterminate Western blots at the first study visit and did not seroconvert to a positive Western blot”

“In December 1999, the FDA notified the Centers for Disease Control and Prevention that blood samples from 91 donors at 26 different blood centers were repeatedly reactive on two or more antibody screening tests (anti-HIV, anti-HTLV-I, and anti-HCV). Additional, more specific tests did not confirm the presence of specific viral antibodies…57 (63%) reported having received the 1991-1992 influenza vaccine prior to their donation and 4 (4%) denied receiving this vaccine. Influenza vaccination history could not be determined for the other 30 donors (33%).”

“Sera from 63.6% of leprosy patients and 23% of their contacts was repeatedly positive by HIV ELISA [which in Africa and England is enough for a diagnosis of HIV infection]”

“False-positive HIV ELISAs have been observed with serum from patients with a variety of medical conditions unrelated to HIV infection [but claims that these can be eliminated by use of Western Blot test or synthetic peptides in the test kits]…False-positive HIV ELISAs [also] occur because of human or technical errors associated with doing the tests or because of antibodies that coincidentally cross-react with HIV or nonviral components in the tests…Notable causes of false-positive reactions have been anti-HLA-DR antibodies that sometimes occur in multiparous women and in multiply transfused patients. Likewise, antibodies to proteins of other viruses have been reported to cross-react with HIV determinants. False-positive HIV ELISAs also have been observed recently in persons who received vaccines for influenza and hepatitis B virus [but again claims that these can be eliminated by Western Blot tests or use of synthetic peptides in tests]”

“[Table shows that of 1,326,030 HIV tests 1,072 had reactive EIA/ELISA tests that were followed by negative or indeterminate Western Blot. Only 276 were confirmed positive]...Some of the EIA repeat reactive [but] Wb [Western Blot] negative or ID [indeterminate] donors were tested on multiple occasions. No donor in...the HIV-1...neg or ID group became Wb positive...The Wb neg or ID donors outnumbered the confirmed reactives 4 to 1 in HIV-1 testing”

“Of the 2,845 repeatedly EIA-reactive eluates [from dried blood samples from newborns] tested by Western blot, 1,323 (47%) were positive, 1,270 (45%) were negative, and 252 (9%) were indeterminate...the number of false-positive Western blots could not be determined in our tested population [they reference a 1989 paper that claimed zero false-positive Western blots by using culture as a validation technique, but this paper reported on only 16 WB+ specimens, and culturing was not performed on WB-indeterminate/negative specimens]”

“Alloimmune mice...were shown to make antibodies against gp120 and p24 of human immunodeficiency virus (HIV), and mice of [two] autoimmune strains...made antibodies against gp120. This is surprising because the mice were not exposed to HIV. [i.e. HIV proteins are found in uninfected mice!!]”

“A common misperception is that, if a serological [antibody] assay is based on recombinant antigens, then results obtained by that assay must, by definition, be completely accurate. In fact, however, certain types of sera...known to generate false-positive results in conventional serological assays can still give false-positive reactions in recombinant antigen-based assays...false-positive reactions have been observed with every single HIV-1 protein, recombinant or authentic.”

“In this study we raised antibodies against Candida albicans [a common infection in immunosuppressed people] mannans of serotype A and B…Applying the Western Blotting techique and a novel enzyme-linked immunosorbent assay [ELISA] system it was established that the antibodies reacted with the gp120 of HIV-1 exclusively…the gp120 protein is exposed on the outer surface of [cultured human] H9 cells where it is recognized by the anti-mannan antibodies”

“Indeterminate patterns can occur in up to 15% of samples tested by WB [Western Blot confirmatory antibody test], and despite recommendations to the contrary, many uninfected persons may be told that they have an indeterminate result without a clear explanation of its meaning. Cumulative experience from more than 5 years of HIV testing suggests that indeterminate WB patterns that persist for many months or years do not indicate HIV infection...37 of 98 indeterminate [blood] donors had WB patterns that were not stable when evaluated over time. In 22 donors whose WB patterns were initially negative, a subsequent WB test showed either single or multiple bands [to proteins believed to be from HIV]. 15 donors whose samples were initially indeterminate became negative at later times. Only 1 of the 98 indeterminate donors showed progression of HIV-1 antibody reactivity to a confirmed positive WB pattern.”

“In blood donor studies in the developed world, about 20% of sera referred to confirmatory laboratories give indeterminate western blot results, almost all of which are on presumed negative specimens.”

“As part of a phase 1 trial of a candidate AIDS vaccine, blood specimens were collected from 168 healthy adult volunteers at minimal or no risk for becoming infected with human immunodeficiency virus type 1 (HIV-1). These specimens were screened for evidence of HIV-1 infection by enzyme immunoassay (EIA) and the Biotech/Du Pont Western blot (168), culture (122), and polymerase chain reaction assay (20). None of the subjects had a positive test result by any of these assays, but 32% had indeterminate Western blot tests, most of which demonstrated a single band of low intensity. The most common bands were p24 (47%), p55 (34%), and p66 (36%); envelope bands were unusual (gp41, 2%; gp120, 2%) [meaning that none of these bands are unique to HIV, yet multiple bands would be interpreted]. No serum specimen collected after 2-11 months from individuals with indeterminate Western blot results was positive by EIA or Western blot. There was 91% agreement in the test results of the first and second serum samples when the same lot of Western blot kit was used but only 36% agreement when different lots were used.”

“Of 168 [healthy, adult] volunteers [for a vaccine trial] tested by WB [Western Blot], 53 (32%) had an indeterminate test result. PBMCs [Peripheral Blood Mononuclear Cells]...from 36 (68%)...were tested for HIV-1 by culture, PCR or both; all were negative...68% had a single [antibody] band; 32% had two or more bands...33 (62%)...returned for follow-up with 2-11 months...Their follow-up serum samples were [still] negative by EIA for HIV-1 p24 antigen and antibodies...Non had developed a positive WB test...Only 12 (36%) of 33 had the same indeterminate WB pattern in both serum specimens; 8 had a different indeterminate WB pattern and 13 had a negative WB results”

“100 ELISA-negative donors...were tested by WB [note that normally a negative ELISA will not result in a Western Blot ‘confirmatory’ test]. 20 were WBi [Western Blot Indeterminate, neither positive nor negative], with p24 being the predominant (70%) and generally the only band. Among recipients of WBi blood, 36% were WBi in their 6 month post-transfusion sample, but so were 42% of a control population that had received only WB-negative blood. When serial samples from recipients with a WB pattern were tested on two occasions, only 35% of results were reproducible. No recipients of WBi blood became ELISA positive, true positive for WB, positive for HIV-1 antigen, or positive for ELISA reactivity against recombinant p24 or gp41. [PCR] was negative for gag and env HIV-1 sequences in all donors and recipients. Thus WBi patterns are exceedingly common in randomly selected donors and recipients”

“A total of 693,000 volunteer blood donors from the Washington, DC, area were screened for HIV infection…from July 1985 through December 1988…1,639 donors tested reactive repeatedly on enzyme immunoassay [ELISA] for HIV, and 284 (17%) of these positive enzyme immunoassays were confirmed by Western Blot [WB] analysis. From this population, 156 donors who tested positive on both [ELISA] and [WB], 64 who tested positive on [ELISA] and negative on [WB], and 16 who tested positive on [ELISA] and whose Western blot results were indeterminate were entered into the study and followed for a median of 28 months…62 of 64 donors (97%) with initially positive [ELISA] but negative [WB] assays became repeated nonreactive on subsequent testing with either the same enzyme immunoassay kit (10%) or the kit of another manufacturer (90%)…46 of 80 donors (58%) selected on the basis of repeatedly reactive [ELISA], but subsequently found to have no physical or laboratory sign of HIV infection, had an indeterminate [WB] pattern during follow-up…In 89% of these cases the bands appeared at a time when the [ELISA] was nonreactive…Blots in 12 donors showed a p24 band alone, in 9 showed p55 alone, and in 5 showed p15 alone. Combinations of bands were seen on the [WBs] of the remaining 20 donors: 10 showed p24 and p55, 4 showed p15 and p55, and 2 showed p15 and p24. Four donors had reactivity with 3 bands - p15, p24, and p55”

“On the basis of these results, it appears that the larger complex [gp160] corresponds to a gp41 tetramer [cluster of four] and the smaller complex [gp120] corresponds to a trimer [cluster of three]. Consistent with this assignment was the occasional detection of a minor band corresponding in size to a dimer. In view of the greater stability and yield of the largest component and the fact that it was the only oligomeric form observed in freshly prepared viral lysates, we propose that the basic quaternay structure of gp41, and consequently of the native HIV surface structure, is a tetramer [i.e. gp160] [If this is true, it means that the detection of gp41, gp120 and gp160 is correlated, and they do not constitute independent evidence of the presence of HIV proteins]”

“[Table 1 shows that out of 630,190 units of blood, 2,119 were positive at least twice on EIA. Out of these, 17 were also positive by Western Blot and culture, 277 had indeterminate WB and 1825 were negative]”

“3 patients who had positive Western blot [antibody] tests are not listed…because later serum samples did not contain HIV-1 antibodies…[in one person this] was felt to be due to passive transfusions of antibody…[the second person] A man who received a heart transplant was HIV-1 seronegative before transplantation. Serum samples taken 16 and 37 days after transplantation were positive for HIV-1 antibodies by EIA; the 37-day sample was also positive for antibodies to p24 and p55 on Western blot testing. 2 subsequent samples obtained 135 and 275 days after transplantation were negative…[the third person] was a 3-year old boy who had a single serum taken 113 days after liver transplantation that was positive by Western blot (p18, p24, p55, p65) but negative for EIA. The serum was tested by Western blot because the EIA result was in the high negative range [EIA/ELISA tests are artificially converted from a continuous test to a black and white answer through the use of an arbitrary cutoff point]. A sample taken 28 days later was negative by both EIA and Western blot. The patient died of multiple bacterial infections 165 days after transplantation [which may have been called AIDS if they had been consistently HIV-positive]”

“We selected the 20 most strongly [indeterminate or atypical Western Blot] reactive samples for further evaluation...Atypical WB [Western Blot] patterns in 19 of 20 of our donors remained substantially the same over time...our data show that the presence of p24 alone in WB should not be regarded as a false positive without subsequent testing of the individual...All study donors had normal immune status...[2] donors were multiparous females [multiple children], and one other probably had received a blood transfusion...we observed a large proportion of individuals who had either lived or worked on dairy farms (6/16) and frequently drank unpasteurized cows’ milk (7/16)..undefined autoimmune phenomena [such as multiple pregnancies], bovine exposure, or cross-reactivity with other human retroviruses could be possible causes for consistently reactive HIV immunologic assays”

“Our data support the use of category I as a positive result [positive WB if 3 of 4 "key" bands are positive - this was the method used by FDA at the time]; however, if this were the only criterion that was finally established as unequivocal positive, more than 50% of patients with AIDS would be put into a ‘probable’ positive or indeterminate category. Our data support the inclusion of categories I, IIa and IIb as unequivocal positives [basically you need 2 of 4 key bands]. Applying these criteria would increase the percent positive for patients with AIDS to 79% without diminishing specificity [but note that 21% of AIDS patients would be HIV-negative!]”

“Natural antibodies capable of neutralizing HTLV-III [HIV] infection of H9 cells were detected in 60% of adult AIDS patients and in 80% of adults with ARC, but in 0% of normal healthy heterosexual controls. Geometric mean antibody titers were two-fold higher in ARC patients compared to AIDS patients and were even higher in 2 antibody positive healthy homosexuals. This finding suggest that virus neutralizing antibodies may exert some in vivo protect effect”

“Most patients (68 to 89%) from low risk groups (prevalence of 0.1% or less) who show reactivity on screening tests will have false-positive results…The predictive value of a positive ELISA varies from 2% to 99%…One notable association with false positive ELISA reactivity in some commercial preparations has been patients with anti-HLA-DR4 antibodies, most often multiparous [having experienced one or more births] or multiply transfused patients...the Western blot method lacks standardization, is cumbersome, and is subjective in interpretation of banding patterns.”

“Although exact percentages were not routinely recorded and calculated[!], approximately 1% of all initial screening ELISA [antibody tests] were reactive [positive], 50% of repeat [second] ELISAs were reactive, and 30% to 40% of first Western blot assays [a more complex antibody test] were reactive and diagnostic [in a very low risk population]”

“Like other viruses, the detection of antibody to HIV provides evidence of past exposure to the virus. Unlike many other viruses, however, the appearance of specific anti-HIV antibodies does not presage clearing of the viremia, loss of infectivity, or clinical recovery. Indeed, anti-HIV seropositive patients, whether asymptomatic or symptomatic, can be assumed to be viremic and current evidence as well as analogy with the biologic features of other retroviruses suggests that most, if not all, such patients will remain so indefinitely [translation: trust us, HIV antibodies are reliable evidence for the presence of the virus. Since ‘HIV’ antibodies were discovered by looking at the blood of people with AIDS, the assumption that antibodies indicate the presence of active virus had already been made]…The predictive value of a positive ELISA varies from 2% to 99% depending on the degree of ELISA reactivity and the presence of risk factors for HIV infection”

“Diagnosis of HIV infection is based almost entirely on detection of antibodies to HIV, but there can be misleading cross-reactions between HIV-1 antigens and antibodies formed against other antigens, and these may lead to false-positive reactions. Thus, it may be impossible to relate an antibody response specifically to HIV-1 infection…Ideally, bands should be seen [on the Western Blot test] at least at p24, p31 and gp41, gp120 or gp160 before a serum specimen is regarded as anti-HIV positive. Indeterminate results in which only one or two bands are seen are not uncommon [proving that no single antigen/antibody reaction is conclusive proof that HIV is present]”

“Inhabitants of certain regions may have cross-reactive antibodies to locall prevalent non-HIV retroviruses”

“Interpretation of Western blots is subjective…these tess have never been submitted to the rigorous evalulations and perfomance assessments under routine laboratory conditions that the screening tests for anti-HIV [e.g. ELISA] have undergone.”

“Monoclonal antibody 5E8 which is specific for a Mr 160,000 glycoprotein (gp160) on the surface of human lung cancer was radiolabeled with 125I. Radiolabeled 5E8 antibody is shown here to suppress the growth of gp160 positive human lung tumor cell lines in a dose-dependent fashion, but this same radiolabeled antibody does not alter the growth of gp160 negative lung tumor cell lines. [gp160 is one of the major 'HIV' proteins, so it is surprising (to believers in HIV antibody tests) to find it without HIV]”

“The strength of ELISA reactivity...was predictive of positivity on immunoblot [Western Blot] testing. The immunoblot was positive in all 21 specimens with ratios higher than 4.0, in 5 of 7 specimens with ratios of 2.0 to 3.9, and in only 1 of 16 specimens with ratios between 1.0 and 1.9 [this illustrates that the HIV test only became a Black and White test through an arbitrary cutoff value]”

“Free HIV antigen and/or low-titre antibodies to recombinant structural or non-structural proteins were seen 6-14 months before seroconversion in all 9 subjects who seroconverted [so, why is the claim made that antibody tests are more than 99% accurate? Why aren’t antigen tests used instead? Could it be because other studies show that HIV antigens aren’t found in about half of HIV-antibody-positive people?]”

“Approximately one-third of AIDS patients in the United States have been from New York City and San Francisco, where, since 1985 [until 1987], less than 7% have been reported with HIV-antibody test results, compared with greater than 60% in other areas”

“Approximately one third of AIDS patients in the United States have been from New York City and San Francisco, where, since 1985, < 7% have been reported with HIV-antibody test results, compared with > 60% in other areas.”

“36 (95%) or 38 African patients with AIDS and 58 (97%) of 60 American patients with AIDS were found to be seropositive to HIV...none of the 100 American heterosexual men was found to be seropositive to HIV, compared with 22 of 100 homosexual American men and six of 100 African outpatients. Common to African patients with AIDS and outpatient controls and American patients with AIDS and homosexual men was the finding of extremely high prevalence rates of antibody to CMV [Cytomegalovirus], HSV [Herpes Simplex], hepatitis B virus, hepatitis A virus, EBV [Epstein-Barr] capsid antigen, syphilis and T gondii. In contrast, the prevalence of antibody to each of these infectious agents was significantly lower among the 100 American heterosexual men”

“Reactivity with [HIV proteins] p24 and/or gp41 has been suggested as a minimum requirement for HIV seropositivity by WB [Western Blot]. While testing ELISA positive serum from Swedish blood donors we detected 3 sera with false-positive WB reactions to p24 and p55...The 3...had no risk factors for HIV infection”

“15,680 donors were screened for [HIV] antibody (Abbott). We found 0.85% initially reactive and 0.37% reactive on at least two of three determinations. The first 38 of these...were sent to laboratories (Abbott) for repeat EIA and confirmatory testing by Western blot. 24 of these samples were from women...with16 indicating on the donor registration card that they had at one time been pregnant...Only 14 of the 38 repeatedly reactive sera...proved to be repeatedly reactive by Abbott. 3 donors (2 men and 1 woman) were Western blot-reactive [i.e. HIV-positive by today’s standards]...the likelihood that a reactive test means previous exposure to HTLV-III [HIV] is low in the blood donor population. It has been estimated, for example, that for a test with 95% sensitivity and specificity, the predictive value ...of a positive test will be only 2%”

“5 of the 220 blood donor specimens [non-reactive on ELISA] were reported to contain anti-HTLV-III [HIV antibodies]...In the high risk group 73 specimens were reported to contain anti-HTLV-III [one believed false positive]...67 reacted against both p24 and gp41, 3 against p24 only and 3 against gp41 only...In this study 6 out of 11 specimens reacting with only one of these two bands [p24, gp41] gave apparently false-positive results [on Western Blot]”

“Seronegative slim disease [African AIDS] patients may represent an immunologically abnormal subgroup of those with the disease or they may have serum antibodies that recognise a variant antigen not detectable by our ELISA assay…or they may have disease unrelated to HTLV-III [HIV] infection…HTLV-III is now endemic (10%) even among symptomless individuals and it may have been so for at least 10 years, since studies on stored sera have shown that a high proportion of these are positive in the ELISA test…[however] like other stored sera coming from a region endemic for malaria, may give a false-positive result on direct binding assay systems, or on western blots.”

“We report the apparent spread of LAV [HIV] in a white American family from husband to wife through heterosexual contact. Although the wife remained clinically well, she developed LAV antibody and a decreased number of T-helper cells. She was followed for 10 months; after exposure to her husband’s semen was discontinued, the LAV antibody was no longer detectable and the T-helper cell number returned to normal [all other family members were consistently negative and clinically well] [This example violates the belief that HIV infections cannot be overcome by the body without drugs, and that HIV antibodies are unambiguous proof of HIV infection]”

“African sera, possibly because they have higher immunoglobulin levels than US and European sera, are highly reactive in ELISA systems and confirmatory assays are essential to rule out false-positive results [but these are often not done in Africa]…6.0% of the 182 persons tested had confirmed HTLV-III [HIV] antibody…The confirmed sera represented 27.5% of those exceeding the screen threshold”

“Serum antibodies to this virus [HIV] have been detected in the majority (68% to 100%) or patients with AIDS and the AIDS-related complex (ARC)”

“If a higher positive cut off (P/N ratio >=4) for the ELISA [antibody test] was used...the number of false-positives would be reduced to 0 of 74 for laboratory and health care personnel, 18 of 1014 for blood donors and 0 of 45 for hemophiliacs. However, this measure would have resulted in several [32] false-negative results [this illustrates that HIV testing is only simplified to Positive versus Negative through an arbitrary cutoff, and this cutoff point is established by its ability to distinguish the majority of people with AIDS from the majority of people who are perfectly healthy, not based on any scientific evidence. It also illustrates that many people have fairly high levels of what are supposedly HIV-specific antibodies, even if they are classified as HIV-negative]”

“Of 49 clinically diagnosed AIDS patients, 43 (88%) showed serum reactivity in this [ELISA antibody] assay…[and were] measured using an ELISA reader which quantifies the color reading. Assays were performed in duplicate; absorbance readings greater than three times the average of 4 normal negative control readings were taken as positive.”

“The sera [from 6720 blood donors] were examined by various enzyme-linked immunoassay (ELISA) screening tests and, usually, by one of three types of confirmatory assay. 45 samples (0.21%) were confirmed as positive. Only 2 were positive in all three confirmatory tests.”

“A confirmed positive test [i.e. one or two ELISA tests, followed by a Western Blot] indicates that a person has been exposed to the virus and has mounted an immunologic response (serum antibodies). However, this test does not indicate whether the person currently harbors the virus”

“When the ELISA is used to screen populations in whom the prevalence of HTLV-III infections is low, the proportion of positive results that are falsely positive will be high”

“Of 96 patients with AIDS or AIDS-related complex and healthy individuals at risk for AIDS, 4 had no detectable antibodies to viral proteins, though [HIV] was isolated from their lymphocytes. 3 of these subjects were symptom-free and 1 had lymphadenopathy. All 4 were sexual partners of patients with AIDS or AIDS-related complex...none of the patients studied here had evidence of impaired production of antibody other viruses”

“Antibody to LAV [HIV] p25 [p24] was found in the serum of 51 of 125 AIDS patients, 81 of 113 patients with lymphadenopathy syndrome, 0 of 70 workers at the Centers for Disease Control (some of whom had handled specimens from AIDS patients), and 0 of 189 random blood donors.”

“A positive [antibody] test for most individuals in populations at greater risk of acquiring AIDS will probably mean that the individual has been infected at some time with [HIV]. Whether the person is currently infected or immune is not known, based on the serologic [antibody] test alone - [HIV] has been isolated [via culture, which is not really isolation] in both the presence and absence of antibody”

“The results provide evidence for the involvement of LAV [HIV] in AIDS...Specific antibodies against [HIV] have been detected in approximately 70% of patients with persistent lymphadenopathy and 40% of AIDS patients studied.”

“antibodies to the structural proteins of HTLV[-I], notably p24 and p19 are not detectable in most AIDS patients [implying that p24 is not part of HIV (HTLV-III)]”

“Clinical samples have also been described have…been described that are reactive in the screening assays but do not contain HIV-1 antibody. Some of these samples possess antibody to certain Class II HLA histocompatibility antigens that are found in some cell lines used to produce the virus. Other persons, who have had no known exposure to HIV-1, produce reactive results in the screening test for still unknown reasons. Such nonspecific results are found commonly when screening tests are used in large populations. Since the psychosocial and medical implications of a positive antibody test may be devastating, it has been recommended that additional testing be performed on such samples [such as this test]…A sample that is reactive in both the EIA screening test and the Western blot is presumed [!] to be positive for antibody to HIV-1, indicating infection with this virus except in situations of passively acquired antibody or experimental vaccination…Sensitivity and specificity of the HIV-1 Western Blot Kit was determined in comparative studies with a previously licensed HIV-1 Western blot using EIA repeatedly reactive samples from high AIDS risk and low risk populations respectively [i.e. without a ‘gold standard’ such as virus purification] [Specificity can only be tested with EIA negative specimens, of which there were none].”