Prospective Evaluation of Amplification-Boosted ELISA for Heat-Denatured p24 Antigen for Diagnosis and Monitoring of Pediatric Human Immunodeficiency Virus Type 1 Infection

The performance in pediatrie human immunodeficiency virus type 1 (HIV-1) infection of a signal-amplification boosted ELISA for HIV-1 p24 antigen in plasma after heat-mediated immune complex dissociation was prospectively compared with polymerase chain reaction-based procedures. Diagnostic sensitivity and specificity of the p24 antigen test were 100% and 99.2%, respectively. Quantification revealed RNA in 85.7% and p24 antigen in 87.4% of 230 samples from 25 infected children. Concentrations of these indices in individual samples correlated (P < .0001). Introduction or modification of antiretroviral treatment showed concordant responses of RNA and p24 antigen in 39 (90.7%) of 43 instances. The treatment-induced changes in concentrations of RNA were higher than those of p24 antigen in 11 instances. In 1 instance, however, the concentration change of p24 antigen was greater than that of RNA (P = .002). Variation of RNA concentrations was more marked than that of p24 antigen (P = .002). The p24 antigen test was equivalent to PCR for diagnosing and monitoring pediatrie HIV-1 infectio

HIV-1 Superinfection in an HIV-2-Infected Woman with Subsequent Control of HIV-1 Plasma Viremia

A human immunodeficiency virus type 2 (HIV-2)-infected woman experienced asymptomatic superinfection with HIV-1 subtype AG. She did not have cross-neutralizing autologous HIV-1 antibodies before and shortly after HIV-1 superinfection. This evidence supports a mechanism other than cross-neutralizing antibodies for the mild course of HIV-1 infection in this woma

Human Immunodeficiency Virus Type 1 p24 Concentration Measured by Boosted ELISA of Heat-Denatured Plasma Correlates with Decline in CD4 Cells, Progression to AIDS, and Survival: Comparison with Viral RNA Measurement

Human immunodeficiency virus type 1 (HIV-1) RNA and p24 antigen concentrations were determined in plasma samples from 169 chronically infected patients (median CD4 cell count, 140 cells/μL; range, 0-1500 cells/μL). p24 quantification involved heat-mediated immune complex dissociation and tyramide signal amplification—boosted ELISA, which has a diagnostic sensitivity similar to that of RNA quantification by a commercial polymerase chain reaction kit. In Cox's proportional hazard models adjusted for CD4 cell count, both RNA (P < .005) and p24 (P = .043) levels were significant predictors of progression to AIDS. Measurement of p24 was superior to measurement of RNA in the model for survival (P = .032 vs. P = .19). p24 level was a significant predictor of CD4 cell decline in models adjusted for CD4 cell counts and was superior or equivalent to RNA level, depending on the group analyzed. Stratification by CD4 cell counts at baseline showed that the superiority of p24 measurement was more pronounced at lower levels of CD4 cells (<200/μL). p24 level may be of interest as a simple and inexpensive predictive marker of disease progressio

Artificial 64-Residue HIV-1 Enhancer-Binding Peptide Is a Potent Inhibitor of Viral Replication in HIV-1-Infected Cells

An artificial HIV-1 enhancer-binding peptide was extended by nine consecutive arginine residues at the C-terminus and by the nuclear localization signal of SV40 large T antigen at the N-terminus. The resulting synthetic 64-residue peptide was found to bind to the two enhancers of the HIV-1 long terminal repeat, cross the plasma membrane and the nuclear envelope of human cells, and suppress the HIV-1 enhancer-controlled expression of a green fluorescent protein reporter gene. Moreover, HIV-1 replication is inhibited by this peptide in HIV-1-infected CEM-GFP cells as revealed by HIV-1 p24 ELISA and real-time RT-PCR of HIV-1 RNA. Rapid uptake of this intracellular stable and inhibitory peptide into the cells implies that this peptide may have the potential to attenuate HIV-1 replication in vivo

Assessment of Recent HIV-1 Infection by a Line Immunoassay for HIV-1/2 Confirmation

Jörg Schüpbach and colleagues show that a second-generation Western blot antibody test used to confirm HIV infection can also be used to determine rates of recent HIV infection

High specificity of line-immunoassay based algorithms for recent HIV-1 infection independent of viral subtype and stage of disease

ABSTRACT: BACKGROUND: Serologic testing algorithms for recent HIV seroconversion (STARHS) provide important information for HIV surveillance. We have shown that a patient's antibody reaction in a confirmatory line immunoassay (INNO-LIATM HIV I/II Score, Innogenetics) provides information on the duration of infection. Here, we sought to further investigate the diagnostic specificity of various Inno-Lia algorithms and to identify factors affecting it. METHODS: Plasma samples of 714 selected patients of the Swiss HIV Cohort Study infected for longer than 12 months and representing all viral clades and stages of chronic HIV-1 infection were tested blindly by Inno-Lia and classified as either incident (up to 12 m) or older infection by 24 different algorithms. Of the total, 524 patients received HAART, 308 had HIV-1 RNA below 50 copies/mL, and 620 were infected by a HIV-1 non-B clade. Using logistic regression analysis we evaluated factors that might affect the specificity of these algorithms. RESULTS: HIV-1 RNA <50 copies/mL was associated with significantly lower reactivity to all five HIV-1 antigens of the Inno-Lia and impaired specificity of most algorithms. Among 412 patients either untreated or with HIV-1 RNA ≥50 copies/mL despite HAART, the median specificity of the algorithms was 96.5% (range 92.0-100%). The only factor that significantly promoted false-incident results in this group was age, with false-incident results increasing by a few percent per additional year. HIV-1 clade, HIV-1 RNA, CD4 percentage, sex, disease stage, and testing modalities exhibited no significance. Results were similar among 190 untreated patients. CONCLUSIONS: The specificity of most Inno-Lia algorithms was high and not affected by HIV-1 variability, advanced disease and other factors promoting false-recent results in other STARHS. Specificity should be good in any group of untreated HIV-1 patients

Cost-Effectiveness of Genotypic Antiretroviral Resistance Testing in HIV-Infected Patients with Treatment Failure

BACKGROUND: Genotypic antiretroviral resistance testing (GRT) in HIV infection with drug resistant virus is recommended to optimize antiretroviral therapy, in particular in patients with virological failure. We estimated the clinical effect, cost and cost-effectiveness of using GRT as compared to expert opinion in patients with antiretroviral treatment failure. METHODS: We developed a mathematical model of HIV disease to describe disease progression in HIV-infected patients with treatment failure and compared the incremental impact of GRT versus expert opinion to guide antiretroviral therapy. The analysis was conducted from the health care (discount rate 4%) and societal (discount rate 2%) perspective. Outcome measures included life-expectancy, quality-adjusted life-expectancy, health care costs, productivity costs and cost-effectiveness in US Dollars per quality-adjusted life-year (QALY) gained. Clinical and economic data were extracted from the large Swiss HIV Cohort Study and clinical trials. RESULTS: Patients whose treatment was optimized with GRT versus expert opinion had an increase in discounted life-expectancy and quality-adjusted life-expectancy of three and two weeks, respectively. Health care costs with and without GRT were $US 421,000 and$US 419,000, leading to an incremental cost-effectiveness ratio of $US 35,000 per QALY gained. In the analysis from the societal perspective, GRT versus expert opinion led to an increase in discounted life-expectancy and quality-adjusted life-expectancy of three and four weeks, respectively. Health care costs with and without GRT were$US 551,000 and $US 549,000, respectively. When productivity changes were included in the analysis, GRT was cost-saving. CONCLUSIONS: GRT for treatment optimization in HIV-infected patients with treatment failure is a cost-effective use of scarce health care resources and beneficial to the society at large

Diagnostic performance of line-immunoassay based algorithms for incident HIV-1 infection

Background: Serologic testing algorithms for recent HIV seroconversion (STARHS) provide important information for HIV surveillance. We have previously demonstrated that a patient's antibody reaction pattern in a confirmatory line immunoassay (INNO-LIA™ HIV I/II Score) provides information on the duration of infection, which is unaffected by clinical, immunological and viral variables. In this report we have set out to determine the diagnostic performance of Inno-Lia algorithms for identifying incident infections in patients with known duration of infection and evaluated the algorithms in annual cohorts of HIV notifications. Methods: Diagnostic sensitivity was determined in 527 treatment-naive patients infected for up to 12 months. Specificity was determined in 740 patients infected for longer than 12 months. Plasma was tested by Inno-Lia and classified as either incident (< = 12 m) or older infection by 26 different algorithms. Incident infection rates (IIR) were calculated based on diagnostic sensitivity and specificity of each algorithm and the rule that the total of incident results is the sum of true-incident and false-incident results, which can be calculated by means of the pre-determined sensitivity and specificity. Results: The 10 best algorithms had a mean raw sensitivity of 59.4% and a mean specificity of 95.1%. Adjustment for overrepresentation of patients in the first quarter year of infection further reduced the sensitivity. In the preferred model, the mean adjusted sensitivity was 37.4%. Application of the 10 best algorithms to four annual cohorts of HIV-1 notifications totalling 2'595 patients yielded a mean IIR of 0.35 in 2005/6 (baseline) and of 0.45, 0.42 and 0.35 in 2008, 2009 and 2010, respectively. The increase between baseline and 2008 and the ensuing decreases were highly significant. Other adjustment models yielded different absolute IIR, although the relative changes between the cohorts were identical for all models Conclusions: The method can be used for comparing IIR in annual cohorts of HIV notifications. The use of several different algorithms in combination, each with its own sensitivity and specificity to detect incident infection, is advisable as this reduces the impact of individual imperfections stemming primarily from relatively low sensitivities and sampling bias

Chapter 32 Human Retroviruses, HIV and HTLV

INTRODUCTION Retroviruses were for many decades well known causative agents of leukemias, lymphomas, other cancers, or chronic inflammations in various animal species. The discovery of the first human retrovirus, human T-cell leukemia virus (now renamed human T-lymphotropic retrovirus type 1; HTLV-1) was reported in 1980 (Poiesz et al., 1981). HTLV-1 was soon identified as the causative agent of adult T-cell leukemia/lymphoma (ATLL), a rapidly progressing cancer of CD4+ T lymphocytes first described in southeastern Japan (Takatsuki et al., 1977). Knowledge gained from HTLV-1 research was important for the subsequent detection of other human retroviruses, first the related HTLV-2 (Kalyanaraman et al., 1982). Soon thereafter, human immunodeficiency virus (HIV)-1 was for the first time isolated from a patient with an early stage of the newly recognized acquired immunodeficiency syndrome (AIDS) (Barre-Sinoussi et al., 1983). Two years later, a second AIDS-causing virus, HIV-2, was discovered (Clavel et al., 1986a). Investigations among nonhuman primates showed a wide distribution of viruses resembling both the HTLV and, respectively, HIV groups of retroviruses. Simian Tlymphotropic retrovirus type (STLV)-1 and STLV-2, simian counterparts of HTLV-1 and HTLV-2, were identified. STLV-3 forms a third group of lymphotropic viruses infecting various African monkey species. HTLV-3, a counterpart of STLV-3 in man, was recently detected in African pygmies (Calattini et al., 2005; Wolfe et al., 2005; Calattini et al., 2006; Switzer et al., 2006). A further HTLV forming a fourth group, HTLV-4, also has been reported (Wolfe et al., 2005). Together, these viruses now constitute four groups of primate T-lymphotropic retroviruses (PTLV-1, -2, -3, -4), with representatives in both simians (STLV) and humans (HTLV). Similarly, both HIV-1 and HIV-2 were shown to originate from primate lentiviruses collectively named simian immunodeficiency viruses (SIV). Other reports of retrovirus infections in humans include isolated cases in which foamy retroviruses (Switzer et al., 2004), simian type-D retrovirus (Lerche et al., 2001) or SIV (Khabbaz et al., 1994) were found in humans as a result of direct cross-species nosocomial transmission from monkeys to caretakers. Transmission of such agents through close and repeated exposure to wild monkeys, for example in bushmeat hunters, also has been reported (Wolfe et al., 2004). Transmission of animal retroviruses to man may not be restricted to viruses of primate origin. Of interest are the recent identification of a betaretrovirus closely related to mouse mammary tumor virus (MMTV) in patients with the autoimmune disease primary biliary cirrhosis (Mason et al., 2004; Xu et al., 2004) and the isolation of an infectious xenotropic murine retrovirus (XMRV) in a form of familial prostate cancer characterized by homozygosity for a reduced activity variant of the antiviral enzyme RNase L (Dong et al., 2007; Fan, 2007). On the other hand, an earlier claim of a novel human retrovirus, has received no follow-up confirmation. This relates to the "human retrovirus 5" (HRV-5), which now has been identified as a rabbit endogenous retrovirus contaminant, RERV-H (Griffiths et al., 2002). An overview of the currently known exogenous human retroviruses and the diseases associated with them is shown in Table 1. In addition, the question whether endogenous human retroviruses might contribute to human autoimmune disease like multiple sclerosis, Sjogren's syndrome, systemic lupus erythematosus and others remains unresolved (Perron et al., 2005; Sander et al., 2005)