THE AIDS INDUSTRY AND MEDIA WANT YOU TO THINK THERE ARE ONLY A HANDFUL OF SCIENTISTS WHO DOUBT THE HIVAIDS THEORY.
HERES THE REALITY.
NAT assays [i.e. viral load] can yield qualitative (i.e., positive or negative) or quantitative (copy number per mL of plasma) results for HIV infection. Methods used for qualitative detection of HIV DNA are laboratory-specific, because no commercially available assays have been approved for clinical use. Methods used to quantitate plasma HIV RNA levels, commonly referred to as viral load tests, utilize a variety of technologies such as conventional and real-time reverse transcriptase (RT)-PCR, as well as amplification methods known as branched chain DNA amplification (bDNA), and nucleic acid sequence-based amplification (NASBA). The intended use of viral load tests is to assess prognosis of disease progression and monitor the effectiveness of ART, rather than diagnosis of HIV infection in HIV-exposed infants [but, despite documenting that neither antibody tests nor NAT tests should be used with infants] The NYSDOH strongly recommends that all New York State birth facilities use the pediatric HIV testing services at the Wadsworth Center. The laboratory performs rapid turnaround NAT in addition to assays capable of detecting HIV-2 infectionDiagnosis of Pediatric HIV Infection in HIV-Exposed Infants. NY State Department of Health. 2010 Nov
http://www.hivguidelines.org/clinical-guidelines/infants-children/diagnosis-of-pediatric-hiv-infection-in-hiv-exposed-infants/Of 1551 HIV-1-infected individuals screened for helminth [parasitic worm] infection, 299 were helminth infected. 234 adults were enrolled and underwent randomization and 208 individuals were included in intent-to-treat analyses. Mean CD4 cell count was 557 cells/microl and mean plasma viral load was 4.75 log10 copies/ml at enrollment. Albendazole therapy resulted in significantly higher CD4 cell counts among individuals with Ascaris lumbricoides infection after 12 weeks of follow-up and a trend for 0.54 log10 lower HIV-1 RNA levels. These effects were not seen with treatment of other species of soil-transmitted helminths. [implying that at least one species of helminth reduces CD4 counts, and that part of what is known as viral load may be due to these parasites]Walson JL et al. Albendazole treatment of HIV-1 and helminth co-infection: a randomized, double-blind, placebo-controlled trial. AIDS. 2008 Aug 20;22(13):1601-9.Data from 1686 women were analyzed: 1203 (71.4%) were categorized as non-users, 429 (25.4%) as intermittent users, and 54 (3.2%) as persistent users of crack [cocaine] Throughout most of the study period, those reporting persistent crack use had higher viral load and poorer immune function, whereas those reporting no use had the lowest HIV-1 RNA levels and best immune health, with intermittent crack users falling in between We found that the median reduction in HIV-1 RNA level was highest in non-users, at 1.7 log10 copies/Ál, compared with 1.4 log10 copies/Ál in inactive crack users and 1.0 log10 copies/Ál in active users. The median CD4 cell count increase was highest in non-users, at 161 cells/Ál, compared with 123 cells/Ál in inactive users, and 100 cells/Ál in active users.Cook JA et al. Crack cocaine, disease progression, and mortality in a multicenter cohort of HIV-1 positive women. AIDS. 2008 Jul 11;22(11):1355-63.PCR results were positive for HIV virus in 3 neonates. These infants on follow up were asymptomatic and 4 have been tested at 18 months using HIV ELISA with two different antigen tests and one rapid test to confirm the diagnosis. Surprisingly, all 3 PCR positive neonates were nonreactive to ELISA.Agarwal D, Agarwal NR. False positive HIV-1 DNA PCR in infancy. Indian Pediatr. 2008;45:245-6.More research is needed to determine the degree to which the viral load in blood predicts the risk of HIV transmission and to determine the association between the viral load in blood and the viral load in semen and vaginal secretionsAntiretroviral therapy and sexual transmission of HIV. UNAIDS. 2008 Feb 1HIV-1 RNA was not prognostic for mortalityPrognosis of HIV-1-infected patients up to 5 years after initiation of HAART: collaborative analysis of prospective studies. AIDS. 2007 May 31;21(9):1185-97.This test is intended for use in conjunction with clinical presentation and other laboratory markers of disease progress for the clinical management of HIV-1 infected patients. The COBAS® AmpliPrep/COBAS® TaqMan® HIV-1 Test is not intended for use as a screening test for the presence of HIV-1 in blood or blood products or as a diagnostic test to confirm the presence of HIV-1 infection. Quantitative measurements of HIV viremia in the peripheral blood have shown that higher virus levels may [!] be correlated with increased risk of clinical progression of HIV disease, and that reductions in plasma virus levels may [!] be associated with decreased risk of clinical progression. Virus levels in the peripheral blood can be quantitated by measurement of the HIV p24 antigen in serum, by quantitative culture of HIV from plasma, or by direct measurement of viral RNA in plasma using nucleic acid amplification or signal amplification technologies. p24 antigen is the principal core protein of HIV and is found in serum either free or bound by anti-p24 antibody. Free p24 antigen can be measured with commercially available enzyme immunoassays (EIA), although the usefulness of p24 antigen as a marker of viral load is limited since the antigen is detectable in only 20% of asymptomatic patients and 40-50% of symptomatic patients the viral protein remains undetectable in most asymptomatic patients. Elevated levels of triglycerides, bilirubin, albumin, hemoglobin and human DNA in specimens as well as the presence of autoimmune diseases such as Systemic Lupus Erythematosus (SLE), Rheumatoid Arthritis (RA) and Antinuclear Antibody (ANA) have been shown not to interfere with the quantitation of HIV-1 RNA or impact the specificity of the COBAS® AmpliPrep/COBAS® TaqMan® HIV-1 Test COBAS AmpliPrep/COBAS TaqMan HIV-1 Test. Roche. 2007 May
http://www.fda.gov/downloads/BiologicsBloodVaccines/BloodBloodProducts/ApprovedProducts/PremarketApprovalsPMAs/ucm092878.pdf +It is advised by the manufacturing companies that viral load tests should not be used as a diagnostic assay, but my belief is that the reason they have done that is there havent been studies done to validate them as diagnostic tests. In practice, however, many people [doctors] do use a viral load test as a test to help confirm the diagnosis of HIV infection. it is just that manufacturers dont recommend it.French MA. Testimony at Parenzee hearing. Court of Criminal Appeal. 2007 Feb[Lawyer] Turner says 'HIV experts acknowledge that there are problems measuring the actual viral load. Different laboratories and different PCR tests obtain markedly different results for the same viral load of the identical specimens'. Do you agree with that?
[Gallo]Yes, I can't testify to the court that it is markedly. There are also variations depending on how they purchase the assay and exactly what you look for and what your primers for the PCR are. We are getting technical but you can look for this part of the genome or you can look for that part of the genome and sometimes these don't exactly correlate one to one. That is true
[Lawyer] Do you agree that the nucleic acid tests, that is the PCR tests, cannot be used to prove HIV infection?
[Gallo] I mean of course you can use it as a component of evidence. You couldn't use it to prove necessarily a virus.Gallo RC. Testimony at appeal hearing of Andre Parenzee. Court of Criminal Appeal. 2007 Feb 12
http://garlan.org/Cases/Parenzee/Gallo.html http://www.bbc.co.uk/worldservice/programmes/worldtoday/news/story/2007/02/070216_kerkhof_shark.shtml South Australian man attacks shark[This article is not about HIV, but it illustrates the dangers of relying on PCR-based tests for any disease] Dr. Brooke Herndon, an internist at Dartmouth-Hitchcock Medical Center, could not stop coughing. For two weeks starting in mid-April last year, she coughed, seemingly nonstop, followed by another week when she coughed sporadically, annoying, she said, everyone who worked with her. Before long, Dr. Kathryn Kirkland, an infectious disease specialist at Dartmouth, had a chilling thought: Could she be seeing the start of a whooping cough epidemic? By late April, other health care workers at the hospital were coughing, and severe, intractable coughing is a whooping cough hallmark It was the start of a bizarre episode at the medical center: the story of the epidemic that wasnt. For months, nearly everyone involved thought the medical center had had a huge whooping cough outbreak, with extensive ramifications. Nearly 1,000 health care workers at the hospital in Lebanon, N.H., were given a preliminary test and furloughed from work until their results were in; 142 people, including Dr. Herndon, were told they appeared to have the disease; and thousands were given antibiotics and a vaccine for protection. Hospital beds were taken out of commission, including some in intensive care. Then, about eight months later, health care workers were dumbfounded to receive an e-mail message from the hospital administration informing them that the whole thing was a false alarm. Not a single case of whooping cough was confirmed with the definitive test, growing the bacterium, Bordetella pertussis, in the laboratory. Instead, it appears the health care workers probably were afflicted with ordinary respiratory diseases like the common cold. Now, as they look back on the episode, epidemiologists and infectious disease specialists say the problem was that they placed too much faith in a quick and highly sensitive molecular test that led them astray At Dartmouth the decision was to use a test, P.C.R., for polymerase chain reaction At Dartmouth, when the first suspect pertussis cases emerged and the P.C.R. test showed pertussis, doctors believed it. The results seem completely consistent with the patients symptoms Then the doctors decided to test people who did not have severe coughing Thats how we ended up with 134 suspect cases, Dr. Kirkland said. And that, she added, was why 1,445 health care workers ended up taking antibiotics and 4,524 health care workers at the hospital, or 72 percent of all the health care workers there, were immunized against whooping cough in a matter of days. If we had stopped there, I think we all would have agreed that we had had an outbreak of pertussis and that we had controlled it, Dr. Kirkland said The Dartmouth doctors sent samples from 27 patients they thought had pertussis to the state health departments and the Centers for Disease Control There was no pertussis in any of the samples when the Centers for Disease Control tested those 39 samples [for antibodies], its scientists reported that only one showed increases in antibody levels indicative of pertussis Dr. Cathy A. Petti, an infectious disease specialist at the University of Utah, said the story had one clear lesson. The big message is that every lab is vulnerable to having false positives, Dr. Petti said. No single test result is absolute and that is even more important with a test result based on P.C.R.Kolata G. Faith in quick test leads to epidemic that wasn't. NY Times. 2007 Jan 22We studied the response to nevirapine-based antiretroviral treatment among women and infants who had previously been randomly assigned to a single, peripartum dose of nevirapine or placebo in a trial in Botswana All women were treated with antenatal zidovudine [AZT]. The primary end point for mothers and infants was virologic failure by the 6-month visit after initiation of antiretroviral treatment By the 6-month visit after the initiation of antiretroviral treatment, 5.0% of the women who had received placebo had virologic failure, as compared with 18.4% of those who had received a single dose of nevirapine Among 60 women starting antiretroviral treatment within 6 months after receiving placebo or a single dose of nevirapine, no women in the placebo group and 41.7% in the nevirapine group had virologic failure. In contrast, virologic failure rates did not differ [statistically] significantly between the placebo group and the nevirapine group among 158 women starting antiretroviral treatment 6 months or more post partum (7.8% and 12.0%, respectively). Thirty infants also began antiretroviral treatment (15 in the placebo group and 15 in the nevirapine group). Virologic failure by the 6-month visit occurred in significantly more infants who had received a single dose of nevirapine than in infants who had received placebo.Lockman S et al. Response to Antiretroviral Therapy after a Single, Peripartum Dose of Nevirapine. N Engl J Med. 2007 Jan 11;356(2):135-147.
http://content.nejm.org/cgi/content/full/356/2/135The patient was a 28-year-old male who was referred for HIV testing after a needle stick injury from a needle contaminated by blood from a newly diagnosed HIV-positive patient. He was given postexposure prophylaxis consisting of zidovudine, lamivudine, and nelfinavir for 4 weeks. Baseline and 6-week HIV-1/2 antibody enzyme immunoassay (EIA) testing produced negative results. An HIV viral load assay was ordered at the same time as the 6-week EIA, for which the patients blood was collected in a Greiner K2 EDTA tube. This tube was centrifuged and then frozen at 70íC within 65 min of collection. The following day, the Versant HIV-1 RNA 3.0 bDNA assay was performed according to the manufacturers instructions, using the Bayer System 340 analyzer, and produced a result of 955 copies/ml. No error code was generated by the analyzer during this test. Immediately after this test was performed, a Bayer technician examined the analyzer by prior arrangement (see below). It was discovered that the read head of the instrument was not adequately compressing the tops of the sample wells, potentially resulting in light leakage between wells. Maintenance was performed to correct this problem. A repeat viral load bDNA assay on the same serum 2 days later produced a result of 75 copies/ml. Additional EIAs performed 3 days and 45 days after the initial viral load assay were repeatedly negative (7 and 12 weeks following the needle stick exposure). An additional viral load assay using new serum collected 2 days after the first viral load also produced a result of 75 copies/ml. In the 4.5 months since the needle stick exposure, the patient continues to be asymptomatic, with negative HIV testing, and is presumed to be HIV negative.Marinovich A et al. False-positive result from a Bayer Versant human immunodeficiency virus type 1 branched-DNA viral load assay, with a possible role for light leakage after inadequate maintenance of the analyzer. J Clin Microbiol. 2006 Nov;44(11):4288-9.A Reactive result indicates that HIV-1 RNA was detected. For a specimen that is repeatedly reactive on an HIV-1 antibody test and reactive in the APTIMA HIV-1 RNA Qualitative Assay, the individual is considered confirmed infected with HIV-1 A specimen that is reactive in the APTIMA HIV-1 RNA Qualitative Assay but that has not been tested in an HIV-1 antibody test should be further tested using a licensed test for HIV-1 antibodies.Aptima HIV-1 RNA qualitative assay. Gen-Probe. 2006 OctPresenting HIV RNA level predicts the rate of CD4 cell decline only minimally in untreated persons. Other factors, as yet undefined, likely drive CD4 cell losses in HIV infection. [Theoretically, HIV RNA represents the amount of virus, and CD4 cells are the type that HIV likes to kill. So there should be a correlation]Rodriguez B et al. Predictive Value of Plasma HIV RNA Level on Rate of CD4 T-Cell Decline in Untreated HIV Infection. JAMA. 2006 Sep 27;296(12):1498-1506.
http://jama.ama-assn.org/cgi/content/full/296/12/1498Clinicians treating patients with HIV encounter some patients with low plasma viral levels who experience rapid progression. What mechanism is responsible for their profound and quick CD4 cell loss? On the other end of the spectrum are those patients with high level HIV viremia who respond clinically like sooty mangabeys infected with simian immunodeficiency virus (SIV), which can tolerate high levels of SIV replication without disease progression. Are such patients statistical extremes in an otherwise simple and uncontested paradigm, or are clinicians and researchers missing something? [like, um, missing something really, really big?] Rodríguez et al have taken the perspective of the individual patient in attempting to quantify how much of the variability of the individual CD4 cell loss is explained by the baseline plasma HIV RNA viral load The provocative main finding from their study was that the presenting plasma HIV RNA load predicted no more than 10% of the observed CD4 cell loss in patients with chronic untreated HIV infection.Henry WK, Tebas P, Lane HC. Explaining, predicting, and treating HIV-associated CD4 cell loss: after 25 years still a puzzle. JAMA. 2006 Sep 27;296(12):1523-5.A malnourished person usually has a high viral load because they have few resources in their body to combat the virus. People with higher viral loads are much more infectious, thus making their partners far more susceptible to getting HIV. Malaria, bilharzia and intestinal worms also drive up a persons viral load.Cullinan K. Radical approach to AIDS prevention. Health-e News (Cape Town). 2006 Jun 5Paradoxically, baseline serum cholesterol, but not atorvastatin, influenced viral rebound at week 4.Negredo E et al. The effect of atorvastatin treatment on HIV-1-infected patients interrupting antiretroviral therapy. AIDS. 2006 Feb 28;20(4):619-21.No correlation was found between DNA proviral load and plasma RNA viral load in thirty consecutive unselected patient samples received for RNA viral load determinations No correlation was found between DNA proviral load and CD4+ cell count in thirty consecutive unselected patient samples received for RNA viral load determinations.Kabamba-Mukadi B et al. Human immunodeficiency virus type 1 (HIV-1) proviral DNA load in purified CD4+ cells by LightCycler real-time PCR. BMC Infect Dis. 2005;5(1):15.A total of 2,937 sera samples, from [Canadian] individuals who were newly diagnosed between 1984 to March 31, 2004 [were examined] Viral RNA was successfully amplified from 2,152 (73.2%) of the sera samples [More than 25% antibody-positive people were negative for viral RNA using the incredibly sensitive PCR test!]HIV-1 strain and primary drug resistance in Canada: Surveillance report to March 31, 2004. Health Canada. 2005 MayHCV [Hepatitis C Virus] has been detected in breast milk and colostrum, and in one study the transmission rate was higher in babies exposed to HCV RNA positive breast milk. However, in most studies no association has been observed between transmission and mode of infant feeding. Of the seven breastfed children in our study with late first positive PCR tests, two were last PCR negative after 4 weeks of age, suggesting possible transmission through breast feeding. However, although we cannot exclude the possibility of postnatal transmission, the occurrence of late last negative [i.e. negative well after birth] and late first positive PCR tests in five of the non-breastfed children as well as in these two breastfed children suggests that these viraemia patterns may not necessarily indicate timing of transmission. We would echo the need to be cautious in inferring the timing of infection simply from early PCR results.Mok J et al. When does mother to child transmission of hepatitis C virus occur?. Arch Dis Child Fetal Neonatal Ed. 2005 Mar;90(2):F156-60.Quantitative PCR should not be used as a diagnostic test for HIV because false positives and false negatives can occur in these circumstancesFearon M. The laboratory diagnosis of HIV infections. Can J Infect Dis Med Microbiol. 2005 Jan;16(1):26-30.several independent contemporary studies have shown that the levels of CSF [cerebral spinal fluid] HIV RNA appear to be significantly lower in untreated subjects with HIV-D [HIV-associated Dementia] than those examined in the era before HAART [indicating that levels of HIV RNA in the CSF is an unreliable surrogate marker]McArthur JC. HIV dementia: an evolving disease. J Neuroimmunol. 2004 Dec;157(1-2):3-10.In our study we had a very high incidence of false positive HIV DNA PCR (75%) especially in younger infantsShah I. Diagnosis of perinatal transmission of HIV-1 infection by HIV DNA PCR. 2004 Oct-Dec;6(4):187-189.Persistent virological response, defined as achieving and maintaining plasma HIV-1 RNA levels at or below the limit of assay quantification (calculated through weeks, 24, 48, and 60), was used as the primary efficacy end pointSaag MS et al. Efficacy and safety of emtricitabine vs stavudine in combination therapy in antiretroviral-naive patients: a randomized trial. JAMA. 2004 Jul 14;292(2):180-9.Test Name: HIV-1 RNA U.Quant/Plasma (>100000 copy/ml). THIS TEST IS NOT FDA APPROVED. FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.Reed S. Test report. UCSD Healthcare. 2004the cumulative proportion of persistent undetectable HIV viral load below 500 copies/ml was significantly higher in the antiretroviral naive patients [those who had never taken AIDS drugs before] than in the non-naive ones In multivariate analysis, being naive of antiretroviral treatment and having a low viral load, at the time of HAART introduction, were significantly correlated with a sustained undetectable HIV viral load.Piroth L et al. Clinical, immunological and virological evolution in patients with CD4 T-cell count above 500/cubic mm: is there a benefit to treat with highly active antiretroviral therapy (HAART)?. Eur J Epidemiol. 2004;19(6):597-604.Survival related significantly to height growth velocity but not to weight growth velocity or head circumference growth velocity. Viral load was not significantly associated with changes in weight-, height-, or head circumference-for-age z scores in children <30 months of age or with changes in weight- or height-for-age z scores in older children.Chantry CJ et al. Growth, survival and viral load in symptomatic childhood human immunodeficiency virus infection. Pediatr Infect Dis J. 2003 Dec;22(12):1033-9.the not infrequent occurrence of borderline quantitative HIV RNA results can confuse the diagnostic status of the child.Parry JV et al. Towards error-free HIV diagnosis: guidelines on laboratory practice. Commun Dis Public Health. 2003 Dec;6(4):334-50.
http://www.hpa.org.uk/cdph/issues/CDPHvol6/No4/6_4guideline1.pdfThe median viral load at the time of diagnosis [of PHI-Primary HIV Infection, a flu-like illness sometimes associated with initial seroconversion] was greater than 750,000 copies/mL (range, 8493-90,397,351 copies/mL [A range of more than 10,000 times!])Weintrob AC et al. Infrequent Diagnosis of Primary Human Immunodeficiency Virus Infection: Missed Opportunities in Acute Care Settings. Arch Intern Med. 2003 Sep 22;163(17):2097-2100.20 HIV-infected subjects were studied A detectable viral load was present in 15 of 17 plasma samples, 10 of 19 BALF [broncho-alveolar lavage fluid] samples, 3 of 20 BF [bronchial fluid] samples, and 1 of 20 saliva samples 42% of subjects who were smokers had detectable viral loads found in BALF samples, [compared to] 71% of nonsmokers Thus, the immunosuppressive properties of smoking may actually inhibit viral replication [more importantly, this study illustrates that the difference between a positive and negative viral load could depend upon the type of sample used to measure it]Wood KL et al. Measurements of HIV viral loads from different levels of the respiratory tract. Chest. 2003 Aug;124(2):536-42.The VERSANT HIV-1 RNA 3.0 Assay (bDNA) is not intended for use as a screening assay for HIV infection or as a diagnostic test to confirm the diagnosis of HIV infection.Summary of Safety and Effectiveness - Versant HIV-1 RNA 3.0 Assay. Versant. 2003 Jul 9From April 1996 through December 2000, a total of 501 antiretroviral-naive [never taken AIDS drugs] HIV-seropositive patients who initiated HAART were recruited at the Hospital Ramón y Cajal [Madrid, Spain] After 24 months of follow-up, 42 (16.5%) of 255 patients were considered to have a discordant immune response [low CD4 cell counts with low viral load or high CD4 cell counts with high viral load] Clinical progression of HIV disease was uncommon among the patients included in the analysis. Overall, 4 patients (1.6%) died of HIV infection-related complications, and 44 patients (17.3%) developed HIV infection-related clinical events Most events (29 [65%] of 44 events) occurred within the first year after initiation of HAART. Overall, clinical events were not more frequent among patients with a discordant immune response than among patients with a good immunologic response.Dronda F et al. Long-term outcomes among antiretroviral-naive human immunodeficiency virus-infected patients with small increases in CD4+ cell counts after successful virologic suppression. Clin Infect Dis. 2002 Oct 15;35(8):1005-9.We report in this work that HIV-1 and HIV-2 patients having a similar degree of CD4 depletion displayed similar levels of T cell hyperactivation and similar numbers of cycling cells in the peripheral blood despite great differences in the plasma viral load. These results and other recent reports call for re-evaluation of different hypotheses about causal relationships among virus concentration, CD4 depletion, and activation and turnover of T lymphocytes.Sousa AE et al. CD4 T Cell Depletion Is Linked Directly to Immune Activation in the Pathogenesis of HIV-1 and HIV-2 but Only Indirectly to the Viral Load. J Immunol. 2002 Sep 15;169:3400-6.56 clinically asymptomatic HIV-1-infected individuals [from Ethiopia], 31 (55%) of whom were also infected with helminths [intestinal worms], were studied At baseline, HIV plasma VL [viral load] was strongly correlated to the number of eggs excreted and was higher in individuals infected with more than one helminth. After treatment of helminths, the 6-month change in HIV plasma VL was significantly different between the successfully treated group and the persistently helminth-positive groupWolday D et al. Treatment of Intestinal Worms Is Associated With Decreased HIV Plasma Viral Load. J Acquir Immune Defic Syndr. 2002 Sep 1;31:56-62.The 2 articles recently published in JAMA are probably among the most important HIV-related papers of the year and have tremendous implications for decision-making. They question the rather arbitrary viral load threshold used in current antiretroviral guidelines and suggest that the main factor that should influence when to start therapy -- and maybe the only one to consider -- should be the CD4+ cell count. The only contributions that viral load can make to this decision are (a) to determine how quickly a given individual is likely to reach the CD4+ cell count threshold at which therapy is indicated, and (b) perhaps to determine the frequency of CD4+ cell-count monitoring. Monitoring viral load only becomes critical when therapy is started.Tebas P. When should antiretroviral therapy be initiated?. Medscape HIV/AIDS. 2002;8(1).The three tests for HIV-1 RNA were 100% sensitive [in this sample of 40] for preseroconversion PHI [i.e. no people diagnosed with Primary HIV Infection symptoms by other methods did not have detectable RNA]. However, bDNA [branched DNA] testing for PHI had a 5% false-positive rate, PCR testing had a 3% false-positive rate and the transcription-mediated amplification assay had a 2% false-positive rate. The false-positive results on the bDNA test ranged from 584 to 2058 copies/ml. False-positive PCR tests ranged from 58 to 103 copies/ml.Hecht FM et al. Use of laboratory tests and clinical symptoms for identification of primary HIV infection. AIDS. 2002 May 24;16(8):1119-29.Prior to the advent of polymerase chain reaction (PCR) technology, physicians relied on HIV culture and p24 antigen testing. Antigen testing is neither sensitive nor specific enough for diagnosis. HIV culture, although accurate, is time-consuming and expensive.Therefore, physicians have quickly embraced PCR technology for testing HIV-exposed infants DNA is preferred [over RNA] for diagnostic purposes because of its sensitivity even in the face of HAART.Benjamin DK Jr. Integration of statistical theory and practical clinical expertise. Polymerase chain reaction testing of the HIV-exposed infant. Minerva Pediatr. 2002 Apr;54(2):105-11.Several potential mechanisms could explain the transient suppression of HIV replication early during the course of measles. Measles virus infection results in lymphopenia, with reductions in CD4+ T lymphocyte percentage and number. The nadir [low point] of the lymphopenia precedes the onset of rash, and values usually return to normal by 1 month after the onset of the rash. This early reduction in CD4+ T lymphocytes could decrease the number of target cells for HIV replication. An alternative explanation is that measles virus infection stimulates the production of soluble factor(s) capable of suppressing HIV replication.Moss WJ et al. Suppression of Human Immunodeficiency Virus Replication during Acute Measles. J Infect Dis. 2002 Mar 25;184.The effect of highly active antiretroviral therapy (HAART) in 85 children infected with human immunodeficiency virus type 1 (HIV-1) was compared retrospectively among Centers for Disease Control and Prevention (CDC) immunologic groups 1-3...HAART resulted in the suppression of HIV-1 below detectable levels in 34.8%, 25%, and 32% of patients in the 3 CDC groups, respectively, and in a frequent switch from syncytium-inducing to nonsyncytium-inducing virus. The results support...the use of other markers of disease progression, in addition to virus load.Nikolic-Djokic D et al. Immunoreconstitution in Children Receiving Highly Active Antiretroviral Therapy Depends on the CD4 Cell Percentage at Baseline. J Infect Dis. 2002 Jan 8;184.The Spearman's rho correlation between viral load in semen and plasma was not significant When restricted to participants with detectable virus in semen and plasma, the correlation remained nonsignificant There was also poor concordance between undetectable viral loads in plasma and semen; 53% of men with undetectable virus in plasma had detectable viral loads in semen and 31% of men with undetectable virus in semen had detectable plasma viral loads Men with equal or greater virus in their plasma than their semen compared with men with greater virus in the semen than plasma showed no differences in age, ethnicity, sexual orientation, years since testing HIV positive, HIV symptoms experienced, CD4+ cell counts, current HIV treatment status, or current HIV treatment adherence [The only correlation found was] that men with greater concentration of HIV in their semen relative to concentrations of HIV in plasma reported significantly greater rates of unprotected vaginal intercourse and total number of unprotected sexual intercourse occasions as the insertive partner Results failed to find differences in STI [sexually transmitted infections] occurrences between men with relatively higher and lower semen viral loads.Kalichman SC et al. Human immunodeficiency virus in semen and plasma: investigation of sexual transmission risk behavioral correlates. AIDS Res Hum Retro. 2001 Dec 10;17(18):1695-1703.[Table 2 shows that the results for identical material sent to multiple laboratories provided viral load results varying from 3,849 to 1,291,635 (Roche Amplicor HIV-1 Monitor), from 63,750 to 205,500 (Bayer HIV-1 3.0 RNA) and from 89,000 to 360,000 (Organon Teknika NucliSens)]Guidelines for Laboratory Test Result Reporting of Human Immunodeficiency Virus Type 1 Ribonucleic Acid Determination. MMWR. 2001 Nov 16;50(RR20):1-12.Available [viral load] tests are not licensed for diagnosing HIV infection, but the viral load test results are used for reporting HIV infection to local and state health departments. Although future versions of these tests might be licensed for diagnostic purposes, healthcare providers should be aware that available viral load tests are only useful for monitoring clinical status after an HIV diagnosis.Guidelines for Laboratory Test Result Reporting of Human Immunodeficiency Virus Type 1 Ribonucleic Acid Determination. MMWR. 2001 Nov 16;50(RR20):1-12.The NucliSens HIV-1 QT assay is not intended to be used as a screening test for HIV-1 nor is it to be used as a diagnostic test to confirm the presence of HIV-1 infection.NucliSens HIV-1 QT. Organon Teknika. 2001 Nov 13
http://davidcrowe.ca/SciHealthEnv/papers/3153-NucliSens-NASBA.pdfAn association was noted between the presence of red blood cells in CVL [Cervico-Vaginal Lavage] and HIV-1 shedding [i.e. detection of RNA believed to be diagnostic for HIV in the cervix/vagina]Kovacs A et al. Determinants of HIV-1 shedding in the genital tract of women. Lancet. 2001 Nov 10;358(9293):1593-1601.Autopsy samples from 13 HIV-infected subjects were examined for HIV-1 viral RNA (vRNA), and viral reverse transcriptase (RT) genotype was determined...The median HIV-1 vRNA level in brain samples from  subjects with moderate dementia was 7.79 log(10) copies/g...compared with 5.44 log(10) copies/g for  mildly demented subjects and 4.87 log(10) copies/g for those obtained from  nondemented individuals...[but] some demented subjects had relatively low levels of HIV-1 vRNA, and paradoxically some nondemented subjects had high vRNA brain levelsMcClernon DR et al. HIV in the brain: RNA levels and patterns of zidovudine resistance. Neurology. 2001 Oct 23;57:1396-1401.HIV-1 RNA was weakly detected in 3 uninfected infants who were negative by DNA PCR at the same age and who ultimately appeared to be RNA undetectable and HIV-1 antibody seronegative [this should be impossible as, if there is no HIV integrated in cells (as DNA) there can't be any virus particles (containing RNA)]Rouet F et al. Early diagnosis of paediatric HIV-1 infection among African breast-fed children using a quantitative plasma HIV RNA assay. AIDS. 2001 Sep 28;15(14):1849-56.That a clinical benefit may not have been achieved with multi-drug rescue therapy calls into question the current wisdom of deeming an undetectable viral load the goal of therapy in the heavily pre-treated population. Even if it can be accepted that an undetectable viral load is an appropriate surrogate marker for clinically relevant outcomes in treatment-inexperienced patients who are initiating combination therapy, it cannot necessarily be accepted without proof that it is a useful surrogate in heavily pre-treated patients...Therefore, until controlled trials are able to prove the utility of an undetectable viral load as a surrogate marker for clinically relevant outcomes in heavily pre-treated patients, we believe that clinicians should show caution before striving for complete viral suppression at any costDeeks SG, Martin JN. Editorial Comment: Reassessing the goal of antiretroviral therapy in the heavily pre-treated HIV-infected patient. AIDS. 2001;15(1):117-9.The difference in HIV RNA between children on ZDV [AZT] and placebo was relatively small. After two years of ZDV, the proportion of IMM children [assigned to AZT at the beginning of the trial, rather than deferred until AIDS developed] with high level (>50%) resistance to ZDV was quite low (38%), and 13% had no ZDV associated mutations [meaning that viral resistance to therapy is not an explanation for anti-HIV therapy to reduce the amount of HIV in the body]Five year follow up of vertically HIV infected children in a randomised double blind controlled trial of immediate versus deferred zidovudine: the PENTA 1 trial. Arch Dis Child. 2001 Mar;84(3):230-6.When it came time to write up the data [on the AIDS 'therapeutic vaccine' called Remune] for publication, Kahn, Lagakos, and others on the team concurred that the analyses showed no benefit from the drug. But scientists from Immune Response performed their own analysis of blood tests on a sample of 250 patients in whom, the company argued, some benefit could be seen - not in longer survival, but in having lower levels of virus ['viral load'] in their blood [Lead researcher] Lagakos said: ''The company did not want our original analysis to go forward. We were put in a position where we had to agree to terms that were unacceptable to us. We decided to go forward with what we had.''Saltus R. AIDS drug researchers say firm pressured them. Boston Globe. 2000 Nov 1Because of the RNA assay's 1.9% to 3.0% false-positive rate, results must be carefully interpreted and compared to HIV-1 viral load levels seen during proven HIV-1 seroconversion. We report the case of a sexually active woman with symptoms suggestive of ARS [Acute Retroviral Syndrome] who had a false-positive HIV-1 RNA assay resultMore D et al. Utility of an HIV-1 RNA assay in the diagnosis of acute retroviral syndrome. S Med J. 2000 Oct;93(10):1004-6.After adjustment, complete nonresponders [CD4 cell counts did not rise, and viral load did not fall after treatment with HAART] and those with only a virologic response had significant higher risks for clinical progression at 6 months [i.e. a reduction in viral load did not reduce the risk for illness]Grabar S et al. Clinical outcome of patients with HIV-1 infection according to immunologic and virologic response after 6 months of highly active antiretroviral therapy. Ann Intern Med. 2000 Sep 19;133(6):401-10.In a small proportion of non-[HIV Type]B samples (about 14%), the HIV RNA copy number, as determined by NASBA, was substantially lower or undetectable compared to results obtained with the other two assays. This phenomenon has also been observed during paediatric diagnosis of HIV In 8 infected African infants and their mothers, although proviral DNA was detected by the Roche Amplicor assay, viral RNA was either not detected or was detected only at a low copy number by NASBA. When infant specimens were tested by the RT-PCR assay, however, HIV RNA was detected and at a higher concentration It has been demonstrated that in some cases although HIV viral load may be undetectable by NASBA, the RT-PCR assay subsequently reports a substantial HIV RNA copy number.OShea S et al. Problems in the interpretation of HIV-1 viral load assays using commercial reagents. J Med Virol. 2000 Jun;61(2):187-94.there was no association [of adherence] with viral load, although only 13 person (31%) met all four adherence goals.Stein MD et al. Adherence to antiretroviral therapy among HIV-infected methadone patients: effect of ongoing illicit drug use. Am J Drug Alcohol Abuse. 2000 May;26(2):195-205.Because viral RNA levels [viral load] are quantified as copies of RNA per milliliter, it is possible that nonviable, but persisting, HIV-1 residues are being detected by such testing. This raises an even more fundamental question as to the validity of viral RNA monitoring in general.Onuigbo MAC. Residual HIV-1 RNA After Highly Active Antiretroviral Therapy. JAMA. 2000 Mar 1;283(9):301-2.Maternal plasma HIV-1 RNA level [viral load] was not associated with perinatal transmission [in this group of women with moderately advanced HIV-1 infection and ZDV/AZT drug resistance]Welles SL et al. HIV-1 genotypic zidovudine drug resistance and the risk of maternal--infant transmission in the Women and Infants Transmission Study. AIDS. 2000 Feb 18;14(3):263-71.Negative results (<50 copies/mL), were obtained in 30/32 (94%) [bDNA 3.0] assays [meaning that false-positive results were obtained in 2/32 or 6% of cases]Erice A et al. Performance characteristics of the bDNA 3.0 assy for quantitation of HIV-1 RNA in plasma [abstract]. 7th Conf. Retroviruses and Opp Infections. 2000 Jan 30-Feb 2Rich and colleagues demonstrated difficulties with the use of HIV-1 plasma viral load testing to diagnose HIV infection. We, too, have witnessed false-positive results on testing for HIV-1 with polymerase chain reaction (PCR) in a man presenting with encephalopathy secondary to alcohol withdrawal and hypertension. A 59-year-old man known to have type 2 diabetes mellitus, alcoholism, hypertension, and chronic obstructive lung disease was hospitalized for headache and confusion. He had no history of trauma, intravenous drug use, sexual promiscuity, homosexual encounters, transfusion, or sexually transmitted disease. Physical examination confirmed disorientation without localizing signs. Magnetic resonance imaging showed increased cerebral atrophy, consistent with the patients age. Lumbar puncture was normal. Nitroprusside was used to control hypertension. The result on enzyme-linked immunosorbent assay for serum HIV was negative, whereas the result on testing for serum HIV-1 RNA by using reverse-transcriptase PCR was positive. Results of repeated enzyme-linked immunosorbent assay of serum and cerebral spinal fluid and PCR assays for HIV RNA were all negative. The HIV-1 PCR assay was designed to monitor HIV therapy, not to diagnose HIV infection. For diagnostic tests, prior probability of a positive test result needs to be considered. In patients (like ours) with a low prior probability of disease, almost all positive test results are false positive. Our urge to confirm the cause of acute encephalopathy rather than accept a diagnosis of exclusion resulted in inappropriate use of HIV-1 PCR. This case confirms the importance of prior probability in diagnostic assays. We concur with Rich and colleagues that lowlevel positive results on HIV-1 PCR must be interpreted cautiously within the context of the patients entire picture.Havlichek DH, Hage-Korban E. False-positive HIV diagnosis by HIV-1 plasma viral load testing. Ann Intern Med. 1999 Nov 16;131(10):794.In contrast to previous reports...the viral load in the majority of the [long-term survivors] tested was detectable and, in some [long-term survivors], quite high...and variable over time.Betts MR et al. Human immunodeficiency virus type 1-specific cytotoxic T lymphocyte activity is inversely correlated with HIV type 1 viral load in HIV type 1-infected long-term survivors. AIDS Res Hum Retroviruses. 1999 Sep 1;15(13):1219-28.The AMPLICOR HIV-1 MONITOR Test is an in vitro nucleic acid amplification test for the quantitation of Human Immunodeficiency Virus Type 1 (HIV-1) RNA in human plasma...[It] is not intended to be used as a screening test for HIV or as a diagnostic test to confirm the presence of HIV infection Quantitative culture has limited utility for monitoring virus levels in infected individuals since only a small fraction of virus particles is infectious in vitro. Infectious virus is often undetectable in asymptomatic individuals The clinical specificity of the test was determined by analysis of 495 anti-HIV-1 negative blood donors. None of these specimens was reactive Assuming[!] a zero prevalence of HIV-1 infection in the seronegative blood donors, the specificity of the test was 100%Amplicor HIV-1 Monitor Test. Roche. 1999Participants with baseline RNA levels below the level defining virological response were excluded from the analysis [which creates an artificial downward trend in the viral load, by eliminating people whose viral load might well go up on therapy]...In the ZDV-naive population [had not taken AZT before this trial] the maximum median fall in HIV RNA occurred by 4 weeks for all three [treatment] arms [AZT, AZT+ddI, AZT+ddC]. Thereafter, the median HIV RNA increased towards baseline levels [so you take these drugs for a lifetime for a theoretical benefit that lasts a month!]... The proportions responding (i.e. achieving HIV RNA [viral load] less than 800 copies/ml) in the first [year] in Delta 1 were 61% on ZDV-ddI and 45% on ZDV-ddC [and 10% on ZDV alone]. In Delta 2 [had previously taken AZT], the proportion of responders [whose viral load dropped] was lower; 23% on ZDV-ddI, and 30% on ZDV-ddC [and 2% on ZDV alone]Delta Coordinating Committee et al. HIV-1 RNA response to antiretroviral treatment in 1280 participants in the Delta Trials: an extended virology study. AIDS. 1999;13:57-65.The results obtained for patients with a broad range of plasma viral loads before and after antiretroviral therapy reveal a constant mean viral (v)RNA copy number (3.6 log10 copies) per infected cell, regardless of plasma virus load or treatment status.Hockett RD et al. Constant Mean Viral Copy Number per Infected Cell in Tissues Regardless of High, Low, or Undetectable Plasma HIV RNA. J Exp Med. 1999 May 17;189(10):1545-54.Compared with patients who maintained undetectable viral load, the adjusted relative hazard of AIDS or death was 1·00 [i.e. no different] for patients with viral rebound [had a viral load that dropped, but later became higher], and 2·40 for patients who failed to reach undetectable concentrations.Ledergerber B et al. Clinical progression and virological failure on highly active antiretroviral therapy in HIV-1 patients: a prospective cohort study. Swiss HIV Cohort Study. Lancet. 1999 Apr 13;353(9156):863-8.18 subjects had 1 or 2 positive results with v.2.0 and an undetectable confirmatory test for a false positive rate of 4.4%. The rate is similar at baseline (9/183 subjects = 4.9%), wk. 4 (7/162= 4.3%) and wk. 26 (2/44 = 4.5%). Of the 18 pos. specimens, 9 tested pos. once and 9 twice. With version 3.0, 11 of 67 samples tested were pos. (16.4%). 6 were pos. once and 5 twice. The range of false pos. rates was 9.1% at wk. 4 (total of 22 specimens) to 26.7% at wk. 26 (total of 15 specimens). A week 4 sample with two values of 8,000 copies/ml on v.2.0 was neg. by DNA PCR, p24 antigen and Western Blot. Follow-up testing of this subject at wk. 26 was negative for HIV antibody and RNA. The emotional impact of a false positive screening RNA test in a recently exposed person is significant. With the high false positive rate, we do not advocate the routine use of HIV RNA tests to screen asymptomatic people. The high rate of repeat false positive tests in a given sample (50%) suggests a possible biologic mechanismRoland ME et al. Pitfalls of HIV RNA testing in the San Francisco post-exposure prevention (PEP) project. Conf Retroviruses Opportunistic Infect. 1999 Jan 31-Feb 4;6(101):Abstract no. 179.Plasma viral load tests for HIV-1 were neither developed nor evaluated for the diagnosis of HIV-1 infection; therefore their diagnostic specificity is not well delineated when applied to persons who are negative for HIV antibody. We report two cases of false-positive results obtained by using branched-chain DNA assay and one case by using HIV reverse transcriptase polymerase chain reaction (RT-PCR) These three cases illustrate the potential problems of using HIV-1 plasma viral load tests for diagnosis of HIV infection Only patients who have a high pre-test probability of a positive result should be evaluated for primary infection by using plasma viral load testing [i.e. preconceptions about risk groups such as gay men makes a positive test more likely] Their performance in patients who are not infected with HIV is unknownRich JD et al. Misdiagnosis of HIV Infection by HIV-1 Plasma Viral Load Testing: A Case Series. Ann Intern Med. 1999 Jan 5;130(1):37-9.We selected 20 healthy volunteers, all of whom yield negative results for HIV antibodies using different screening tests. Plasma from all of them were analysed by three different currently available HIV viral load tests: branched DNA (bDNA) signal amplification assay (Chiron), nucleic acid sequence-based amplification (NASBA) Nuclisens (Organon Teknika), and Ultradirect reverse transcriptase (RT)-PCR Monitor (Roche) 2 samples [10%] yielded positive results by the bDNA assay Another 2 specimens [10%] yielded false-positive results by the NASBA Nuclisens [and] one of the 20 samples [5%] was interpreted as positive by the Ultradirect RT-PCR Monitor assay using the Monitor test with non-B primers, up to 4 of the 20 samples [20%] yielded positive values Results were reproduced in more than half of tested specimens for which plasma volumes were enough for repeat testingde Mendoza C, Holquin A, Soriano V. False positives for HIV using commercial viral load quantification assays. AIDS. 1998;12(15):2076-7.A reanalysis is presented of 43 cases of apparent transient viremia [one or more positive culture or PCR assays for HIV-1 and the subsequent inability to detect HIV-1 in the specimens on multiple occasions or seroreversion, or both]. 41 cases occurred among 1561 infants in five studies of mother-to-infant HIV-1 transmission...Our negative studies of 43 cases of suspected transient infection indicate that the phenomenon...remains to be proven and that most cases suggestive of transient HIV-1 infection are cases of mislabeling of specimens or their contamination in the laboratory...Frenkel LM et al. Genetic evaluation of suspected cases of transient HIV-1 infection of infants. Science. 1998 May 15;280(5366):1073-7.Specimens were studied from one mother and her child, both with suspected transient viremia. The mother had two and the infant three positive HIV-1 cultures, but subsequently both individuals became negative for HIV-1 by nPCR, standard virus cultures, CD8+-depleted virus cultures, and enzyme-linked immunosorbent assay. HIV-1 RNA and DNA were not detected in two lymph nodes taken from the mother 3 and 4 years after the last virus-positive culture. PCR amplification and DNA sequencing of HIV-1 env sequences from the mothers two and the childs three culture supernatants were performed in separate laboratories to eliminate the possibility of cross-contamination. Phylogenetic analysis found that none of the five isolates were genetically linked. Although it is improbable, these five virus isolates appear to have arisen from five separate incidents of specimen contamination or mislabeling. This case remains enigmatic, however, in that both the mother and infant had strong CD8+ cytotoxic T lymphocyte (CTL) responses and lymphocyte proliferation to multiple HIV-1 antigensFrenkel LM et al. Genetic evaluation of suspected cases of transient HIV-1 infection of infants. Science. 1998 May 15;280(5366):1073-7.[This study was a] Prospective follow-up of a cohort of 162 unselected, protease inhibitor-naive, antiretroviral-experienced patients with advanced HIV disease, treated with indinavir combined with two nucleoside analogues 21% of patients exhibited discrepant virological and immunological responses to treatment, of whom one-half failed to exhibit significant increases in CD4 cells despite a virological response to therapy and one-half exhibited increased CD4 cell counts in the absence of significant decrease in plasma viral load. The incidence of AIDS-defining events in the latter group of patients was similar to that of responder patients, whereas their incidence was higher in patients who failed to exhibit a virological and immunological response and those who failed to increase CD4 cells despite a significant decrease in viral load. [calling into question the common use of viral load to estimate the rate of progression to AIDS]Piketty C et al. Discrepant responses to triple combination antiretroviral therapy in advanced HIV disease. AIDS. 1998 May 7;12(7):745-50.Peak levels [of HIV-1 RNA] in the first 120 days were not predictive of disease progressionSchacker TW et al. Biological and Virologic Characteristics of Primary HIV Infection. Ann Intern Med. 1998 Apr 15;128:613-20.
http://www.annals.org/cgi/content/full/128/8/613the use of absolute viral levels is problematic for individual patient management because the assessment of the absolute viral RNA level is dependent upon the test method.Brambilla D, Leung S, Lew J et al. Absolute copy number and relative change in determinations of human immunodeficiency virus type 1 RNA in plasma: effect of an external standard on kit comparisons. J Clin Microbiol. 1998 Jan;36(1):311-4.Unspecific pick-ups during PCR can be due to annealing of primers to target sequences with a low degree of homologyDennin RH, Chen Z. Hepatitis C virus (HCV) specific sequences are demonstrable in the DNA fraction of peripheral blood mononuclear cells from healthy, anti-HCV antibody-negative individuals and cell lines of human origin. Eur J Clin Chem Clin Biochem. 1997 Dec;35(12):899-905.Initial testing of plasma by RNA RT-PCR was reported as positive. This result was not confirmed by viral cultures, nested DNA PCR, western blot, or EIA. Additional RNA RT-PCR assays gave positive results from earlier occasions in the vaccine trial. Eventually, testing of all previously reactive samples by RNA RT-PCR was repeated in a quality-controlled laboratory, and confirmed the negative HIV-1 status of the individual. Interpretation This case report exemplifies the difficulties with use of viral-genome measurement as a screening test to diagnose HIV- 1 infection, particularly in individuals who have ever participated in HIV-1 vaccine trials. Monitoring of large numbers of phase III vaccinees by RNA RT-PCR will not be feasible. The design of efficacy trials for new vaccines should be in parallel with development of antibody-based diagnostic tests that are capable of differentiating between immunisation and true HIV-1 infection.Schwartz DH et al. Extensive evaluation of a seronegative participant in an HIV-1 vaccine trial as a result of false-positive PCR. Lancet. 1997 Jul 26;350(9073):256-9.[In this study of 78 HIV-positive people with high CD4 cell counts and no symptoms] there were no differences in viral load with regard to time of HIV-1 infection [i.e. amount of virus does not grow over time] 10 patients fulfilled the criteria for LTNP [Long Term Non-Progressors]. 7 of these 10 patients had viral loads above 10,000 RNA copies/ml and 2 above 30,000 RNA copies/ml. The level of viral load of LTNP was not statistically different compared with the other 68 patientsGarcía F et al. Viral load in asymptomatic patients with CD4+ lymphocyte counts above 500 million/l. AIDS. 1997;11:53-7.We observed that clade A strains [of HIV] were not detected by RT-PCR [Reverse Transcriptase-Polymerase Chain Reaction] and that clade G-strains were not detected by NASBA [Nucleic Acid Sequence-Based Amplification]. However, the copy number detected by RT-PCR in one clade E (CM235) and in one clade F (163.3070) was much lower than the copy number detected by bDNA [branched DNA] and NASBA...for clades B and D (UG270), the HIV-1 RNA levels measured by bDNA were lower than those obtained by RT-PCR and NASBACoste J et al. Effect of HIV-1 genetic diversity on HIV-1 RNA quantification in plasma: comparative evaluation of three commercial assays. J Acquir Immune Defic Syndr. 1997 Jun 1;15(2):174-5.The copy number obtained with the bDNA assay tended to be lower than that obtained by the reverse-transcription-PCR assay...by a median factor of two.Sperling RS et al. Maternal viral load, zidovudine treatment and the risk of transmission of human immunodeficiency virus type 1 from mother to infant. N Engl J Med. 1996 Nov 28;335(22):1621-9.Specimens were tested twice with PCR; only those for which PCR reaction was repeatedly positive were reported as positive [which indicates that false positives occurred at a significant level, although no numbers were reported. Furthermore, if a false positive can occur once, it can occur twice]Bertolli J et al. Estimating the timing of mother-to-child transmission of human immunodeficiency virus in a breast-feeding population in Kinshasa, Zaire. J Infect Dis. 1996 Oct;174(4):722-726.for 223 specimens for HIV-infected infants [based on two positive cultures], 89% had PCR positivity, 11% had PCR negativity and <1% had indeterminate status.Bremer JM et al. Diagnosis of infection with human immunodeficiency virus type 1 by a DNA polymerase chain reaction assay among infants enrolled in the women and infant's transmission study. J Pediatr. 1996 Aug;129(2):198-207.maintenance of plasma HIV RNA levels below 10,000/ml in early HIV disease appears to be associated with decreased risk of progression to AIDS. However, in patients with more advanced disease [low CD4 cell counts], disease progression occurred in up to 30% of patients with fewer than 10,000 HIV RNA copies/ml [In other words, viral load is not terribly well associated with progression to AIDS]...HIV RNA levels should not be measured within a month of acute illnesses or within a month after influenza and pneumococcus immunizations. Increases in HIV RNA levels in blood of as much as 300-fold have been observed within two weeks of routine immunizations against influenza, tetanus, or pneumococcusSaag MS et al. HIV Viral load markers in clinical practice. Nat Med. 1996 Jun;2(6):625-9.Our investigation produced two main findings. First, the false-positive and false-negative rates of PCR that we determined are too high to warrant a broader role for PCR in either routine screening or in the confirmation of diagnosis of HIV infection. This conclusion is true even for the results reported from more recent, high-quality studies that used commercially available, standardized PCR assays...We did not find evidence that the performance of PCR improved over timeOwens DK et al. Polymerase chain reaction for the diagnosis of HIV infection in adults. A meta-analysis with recommendations for clinical practice and study design. Ann Intern Med. 1996;124:803-15.Testing for HIV by PCR or culture also may be helpful in determining HIV status; however, neither test is licensed for diagnosis of HIV infection.US Public Health Service guidelines for testing and counseling blood and plasma donors for Human Immunodeficiency Virus Type 1 antigen. MMWR. 1996 Mar 1;45(RR-2).the majority (83%) of [flu] vaccinated [HIV-positive] individuals experienced a significant increase in plasma HIV-1 RNA levels within 1-2 [weeks after] immunization and returned to their prevaccination levels within 4 weeks after immunization...patients on antiretroviral therapy were not noticeably different from those not in therapy with regard to increases in plasma viremiaStaprans SI et al. Activation of virus replication after vaccination of HIV-1 infected individuals. J Exp Med. 1995 Dec 1;182(6):1727-37.The sensitivity of the polymerase chain reaction [PCR] technique has been estimated at 30-50% during the first week of life, 70-90% at 1-2 months of age and >95% after 3 months of age However, the techniques used are not standardized, there is no common quality control programme, few results are available from prospective cohort studies in developing countries and the detection rates in the neonatal period are lowDabis F et al. Methodology of intervention trials to reduce mother-to-child transmission of HIV-1 with special reference to developing countries. AIDS. 1995;9(Suppl A):67-74.TABLE 1. Comparison of the estimated mean number of RNA copies per milliliter of plasma with the mean titer of virus in plasma and the p24 antigen level in symptomatic and asymptomatic patients [for symptomatic, HIV-positive patients 100% had detectable RNA viral load but only 34/41 of these had positive virus cultures and only 22/35 had p24 antigen detectable] [for asymptomatic HIV-positive patients 29/39 had viral load detectable at low levels but only 5/39 had positive virus cultures and 5/38 detectable p24 antigen]Van Kerckhoven I et al. Quantification of human immunodeficiency virus in plasma by RNA PCR, viral culture, and p24 antigen detection. J Clin Microbiol. 1994 Jul;32(7):1669-73.the high level of plasma virus observed by Piatak et al, was about 99.9 per cent non-culturable, suggesting that it was either neutralized or defective.Sheppard HW, Ascher MS, Krowka JF. Viral burden and HIV disease. Nature. 1993 Jul 22;364(6435):291-2.96% (105 of 110) of all [HIV-positive] patients tested had quantifiable plasma RNA [although that is high, one wonders how the others can have HIV in their bodies without detectable amounts of HIV RNA (free HIV particles) using PCR, an extraordinarily sensitive method]Winters MA et al. Biological variation and quality control of plasma Human Immunodeficiency Virus type 1 RNA quantitation by reverse transcriptase Polymerase Chain Reaction. J Clin Microbiol. 1993;31(11):2960-6.Primary infection with [HIV] is generally followed by a burst of viraemia [high 'viral load'] with or without clinical symptoms. This in turn is followed by a prolonged period of clinical latency. During this period there is little, if any, detectable viraemia, the numbers of infected cells in the blood are very low, and it is extremely difficult to demonstrate virus expression in these cells in patient 1 [high CD4 cell count], the frequency of HIV-infected cells is 1/10,000 in the PB [Peripheral Blood], and between 1/100 and 1/1,000 in the LN [Lymph Nodes]. In patient 6 [moderately low CD4 cell count], the frequency is between 1/1,000 and 1/10,000 in the PB, and 1/1000 in the LN. In patient 10 [low CD4 cell count], the frequency is between 1/10 and 1/100 in both PB and LN [but note that Piatak (1993) showed evidence that only 1/60,000 infected cells were actually viable, infectious virions, and there is no word in the paper of which, if any, of these 12 people studied were ill]Pantaleo G et al. HIV infection is active and progressive in lymphoid tissue during the clinically latent stage of disease. Nature. 1993 Mar 25;362(6418):355-8.Most CD4+ lymphocytes must be latently infected because we did not detect viral RNA in more than a small fraction of the cells in which HIV DNA was detectable. One in 100-400 cells with HIV DNA had viral RNA as abundant as productively infected cells; and a higher proportion of cells had low levels of viral RNA [up to 6%]Embretson J et al. Massive covert infection of helper T lymphocytes and macrophages by HIV during the incubation period of AIDS. Nature. 1993 Mar 25;362(6418):359-62.Circulating levels of plasma virus determined by QC-PCR also correlated with, but exceeded by an average of nearly 60,000-fold...titers [amounts] of infectious HIV-1 determined by quantitative endpoint dilution culture of identical portions of plasma.Piatak M Jr et al. High levels of HIV-1 in plasma during all stages of infection determined by competitive PCR. Science. 1993 Mar 19;259:1749-54.Our study of PBMC [Peripheral Blood Mononuclear Cells] from 56 HIV-1-seropositive patients, using in situ hybridization alone, also [as did other studies] revealed only 1 in 5,000 to 1 in 100,000 cells positive for HIV-1-specific nucleic acids [DNA]...Our finding with the use of in situ PCR that large numbers [if you consider 0.1% to 13.5% of cells a large number] of PBMC from HIV-1-seropositive patients contain the provirus suggests that direct cytopathic [cell-killing] effects of the virus may be an important but not necessarily the sole cause of depletion of CD4-positive lymphocytes. Our data also argue strongly against the theory that HIV-1 is not the primary etiologic [causative] agent of AIDS [huh?]...we were able to...show a relation between viral load and the stage of HIV-1 clinical infection. Patients in Stage II [HIV+, but no symptoms] had a significantly lower percentage of HIV-1-positive PBMC than those in Stage III and Stages IV-A to IV-C [but the authors neglect to mention that this is only true of the average value, there was considerable overlap between individual values. More importantly, they also do not mention that the average value at Stage III (lymph gland enlargement) was higher than at Stage IV-A to C (AIDS) and much higher than at Stage IV-D (Kaposis Sarcoma)]. Patients in Stage IV-D (who had Kaposis sarcoma only) had relatively low numbers of HIV-1-infected cells.Bagasra O et al. Detection of human immunodeficiency virus type 1 provirus in mononuclear cells by in situ polymerase chain reaction. N Engl J Med. 1992;326(21):1385-91.This proficiency study of PCR detection of HIV-1 DNA in serum identified a disturbingly high rate of nonspecific positivity with a widely employed gag primer pair system. In fact, the overall rate of positivity was not significantly different for serum specimens from seropositive patients and seronegative control donors (26% versus 18%)Busch MP et al. Poor sensitivity, specificity, and reproducibility of detection of HIV-1 DNA in serum by polymerase chain reaction (PCR). J Acquir Immune Defic Syndr. 1992;5(9):872-879.PCR performed before the age of 1 month identified only 55% of the truly infected infantsVan de Perre P et al. Postnatal transmission of Human Immunodeficiency Virus 1 from mother to infant: a prospective cohort study in Kigali, Rwanda. N Engl J Med. 1991;325:593-8.The [HIV-1 DNA PCR, viral load] assay did not appear more sensitive than classic serologic tests in the diagnosis of HIV-1 infection [all seronegative individuals were negative on DNA PCR, including those who had had unprotected sex with seropositive people]...it was even less sensitive, because the specimen from one of our seropositive individuals was negative on PCR with the three primer pairs. Our results are consistent with...Jackson, Taylor, and Lifson...However, our results are not consistent with those of other studies: Loche, Wolinski, Imagawa, and Ameisen observed positive PCRs in seronegative at-risk individuals. These discrepancies are difficult to explain.Lefrere JJ et al. No evidence of frequent HIV-1 infection in seronegative at-risk individuals. Transfusion. 1991;31(3):205-11.The DNA equivalent of 200 microliters of blood was assayed using the pol, gag, and nef primer pairs in a single PCR, and by performing a nested gag PCR [on] 11 Western blot anti-p24(gag)-indeterminate blood donors. No positive signal was found for any of these donors.Bruisten SM et al. Enhanced detection of HIV-1 sequences using polymerase chain reaction and a liquid hybridization technique. Application for individuals with questionable HIV-1 infection. Vox Sang. 1991;61:24-9.false positive PCR results were unrelated to the childs age [i.e. occurred at all ages up to 18 months]. PCR was recently proposed as a diagnostic method for HIV-1 infection in neonates in order to initiate early specific therapy. Our findings suggest the need for caution in using this test alone as evidence of true HIV-1 infection. In addition, positive findings with at least two primer pairs in the sample do not avoid the possibility of sample contamination by exogenous DNA and, in our opinion, positive results in at least two different subsequent samples are required.De Rossi A et al. Antigen detection, virus culture, polymerase chain reaction, and in vitro antibody production in the diagnosis of vertically transmitted HIV-1 infection. AIDS. 1991 Jan;5(1):15-20.Only 50% of collaborating AIDS Clinical Trial Group laboratories were able to detect 60 copies of HIV DNA in 150,000 cells in a National Institutes of Health PCR standardization programme. 33% of these laboratories had problems with false positives.Tudor-Williams G. Early diagnosis of vertically acquired HIV-1 infection. AIDS. 1991 Jan;5(1):103-5.Among the 8 newborns born to HIV1-infected mothers, 6 had detectable HIV1 DNA sequences In group II [babies], 17 of 23 sera were available HIV1 DNA was detected in PBL [peripheral blood leukocytes] of 14 of 23 babies. Among the 14 babies positive by PCR, 10 had positive or incomplete and 2 had negative Western blots. Among the nine babies negative by PCR, 3 had positive or incomplete and two had negative Western blots. Therefore, 2 of 4 babies, who were seronegative at the time of PCR test, had HIV1 DNA in PBL 8 (including the 4 with symptoms consistent with AIDS) of the 14 HIV1 DNA-positive and 1 of 9 HIV DNA-negative babies had clinical features likely to be related to HIV infection. The results obtained with the three sets of primers markedly differed; indeed 7 of 31 samples yielded positive results with all three sets of primers as compared to 7 of 31 which were positive with two sets of primers but negative with the third, and the remaining six of 31 samples were only positive with one set.Paterlini P et al. Polymerase chain reaction for studies of mother to child transmission of HIV1 in Africa. J Med Virol. 1990 Jan;30(1):53-7.We found that in infected but asymptomatic patients with HIV-1, [an average of] 1 in 50,000 PBMC harbored the virus. When such a patients condition progressed to AIDS-related complex or AIDS, the viral titer increased significantly, to approximately 1 in 400 PBMC...It is unclear from this study, however, what percentage of the infected cells carry HIV-1 latently and what percentage of the cells express the virus actively. If 1 in 10,000 PBMC...express viral messenger RNA...then 99.6% of the infected mononuclear cells harbor the virus latently and the remaining 0.4 % express it actively [i.e. 1 in 100,000 cells have active virus!]...this information on the quantitation of HIV-1 [high levels of HIV-1 viremia] should reduce residual doubts about whether HIV-1 is the true etiologic agent of AIDSHo DD, Moudgil T, Alam M. Quantitation of Human Immunodeficiency Virus type 1 in the blood of infected persons. N Engl J Med. 1989 Dec 14;321(24):1621-5.17 of 18 DNA samples from seropositive subjects (94%) had positive [PCR] signals 13 of 21 (62%) RNA samples from seropositive subjects were positive for HIV RNA RNA samples also gave varying intensities of PCR signal.Hart C et al. Direct detection of HIV RNA expression in seropositive subjects. Lancet. 1988 Sep 10;2:596-9.we analyzed samples from individuals known to be at especially high risk of HIV infection-seronegative sexual partners of seropositive individuals...Of 16 seronegative partners tested, 5 were unequivocally positive for HIV DNA. The clinical records of these 5 subjects confirmed that they were seronegative by enzyme-linked immunosorbent assay and western blot and negative for the p24 antigen at the time the blood samples were taken for the DNA assay...the same 5 samples were found to be positive with a second HIV-specific oligonucleotide...The serological [antibody] status was confirmed in each case and each of the 5 individuals was negative for anti-HIV antibodies and p24 antigen 2 and 3 months after the initial detection of HIV DNALoche M, Mach B. Identification of HIV-infected seronegative individuals by a direct diagnostic test based on hybridisation to amplified viral DNA. Lancet. 1988 Aug 20;2(8608):418-21.Testing for HIV by PCR [viral load] or culture also may be helpful in determining HIV status; however, neither test is licensed for diagnosis of HIV infectionCDC. Public Health Service Guidelines for Counseling and Antibody Testing to Prevent HIV Infection and AIDS. MMWR. 1987 Aug 14;36(31):509-15.
http://www.cdc.gov/mmwr/PDF/RR/RR4502.pdfDetailed characterization of HTLV-III [now called HIV] and serologic [antibody] testing of large numbers of patients with AIDS or AIDS-related complex (ARC) became possible when it was found that the virus could be transmitted to a human T-cell line, H9, that is largely resistant to the cytopathic effects of the virus but is a good virus producer Sequences of HTLV-III were first detected in DNA of infected H9 cells by Southern blot analysis There was no evidence of such sequences in uninfected H9 cells [but] In all, HTLV-III sequences were detected in only 9 out of 65 patients [with AIDS or ARC] evaluated. Roughly the same portion of ARC patients (3 out of 26) as AIDS patients (6 out of 39) was positive for HTLV-III None of five Kaposi's sarcoma tissue specimens examined was positive for HTLV-III DNA sequences Southern hybridizations can detect less than one viral DNA copy per 10 cells, [therefore] we interpreted these data to indicate that the three Kaposi's sarcoma lesions did not contain HTLV-III sequences In most of the patients in which HTLV-III DNA sequences were detected in fresh tissue, the signal intensities were weak [estimated as] at a level of less than one copyy per 10 cells Unlike most other retroviruses, HTLV-III appears to persist in both integrated [DNA within the nucleus] and unintegrated [DNA floating around in the cell unattached] forms in chronically infected cells.Shaw GM et al. Molecular characterization of human T-cell leukemia (lymphotropic) virus type III in the acquired immune deficiency syndrome. Science. 1984 Dec 7;226(4679):1165-71.
There you have it. No handful of wild-eyed conspiracy theorists. No right-wing racists, as the Aids industrys spinmeisters would have you believe. Just 19 very serious, concerned, highly educated people from every corner of the globe who sense that an enormous tragedy is unfolding due to the medical establishments unwillingness to face the evidence that the Hiv-Aids theory is a mistake.
The people on this page were intellectually curious enough to have sought out and studied the arguments that discredit the Hiv-Aids theory. Since the mass media and professional journals censor these arguments, the vast majority of doctors and scientists, although decent people who want to do the right thing, have never been exposed to them, and so accept the biased conclusions of politicized bureaucracies like the CDC and WHO, whose coziness with the drug industry is legendary and whose recommendations always seems to dovetail perfectly with drug industry marketing plans.
Were it not for the massive media blackout of information that contradicts the Hiv theory, many more people would be asking tough questions.
The next time you hear the media say, only a handful of scientists doubt Hivs role in Aids, refer them to this page. Explain to them that it is wrong to misrepresent the fact that there is enormous dissent to the Hiv-Aids paradigm.
The next time you hear the media drone, Hiv, the virus that causes Aids, remind them that journalists are supposed to distinguish between what is a theory and what is a fact. That Hiv-Aids is only a theory and has never been proven, is admitted by top scientists even in the Aids establishment.
The next time the media announce that tens of millions of people are dying from Hiv in Africa, ask them how they know that. Remind them that journalists are supposed to question dubious assertions from powerful, drug-industry funded agencies like the WHO, not parrot them as if they were indisputable. Ask them why they report these numbers as if they were actual Aids cases, when in fact they are projections made by WHOs computer programs, based on very questionable statistical methodologies and contradicted by many facts including the continual large population increases experienced in the countries supposedly worst affected.
Request that the media stop twisting the truth in support of a politicized, entrenched Aids establishment that profits financially by terrorizing people, pokes its nose shamelessly into peoples private sex lives, compels people to submit to inaccurate tests and literally forces mothers and babies to swallow toxic, unproven chemotherapy drugs with horrific, often-fatal side effects.
Explain to them that this is irresponsible, and that such actions cause needless anxiety, shatter peoples lives, tear families apart, destroy hope and trigger countless suicides. And that while we realize that sensational headlines about killer viruses sell newspapers, the social cost of these profits is unacceptable.
Make the media understand that keeping people in the dark about the large number of credentialed dissenters to the Hiv-Aids dogmas, and the financial conflicts of interest that are rampant among Hiv-Aids scientists and NGOs, is a violation of everyones human right to informed consent and freedom of information.
Note: Affiliation with an organization does not imply that the organization supports the individual HIV/AIDS skeptics position.
Does Hiv cause Aids? Lots of scientists say no. Read more.
Alive And Well
Dr. Peter Duesberg
The Perth Group
Treatment Information Group
Immunity Resource Foundation
Alberta Reappraising Aids Society
Last updated January 29, 2013.