Assessment of recent HIV-1 infection by a line immunoassay for HIV-1/2 confirmation.

BACKGROUND: Knowledge of the number of recent HIV infections is important for epidemiologic surveillance. Over the past decade approaches have been developed to estimate this number by testing HIV-seropositive specimens with assays that discriminate the lower concentration and avidity of HIV antibodies in early infection. We have investigated whether this "recency" information can also be gained from an HIV confirmatory assay. METHODS AND FINDINGS: The ability of a line immunoassay (INNO-LIA HIV I/II Score, Innogenetics) to distinguish recent from older HIV-1 infection was evaluated in comparison with the Calypte HIV-1 BED Incidence enzyme immunoassay (BED-EIA). Both tests were conducted prospectively in all HIV infections newly diagnosed in Switzerland from July 2005 to June 2006. Clinical and laboratory information indicative of recent or older infection was obtained from physicians at the time of HIV diagnosis and used as the reference standard. BED-EIA and various recency algorithms utilizing the antibody reaction to INNO-LIA's five HIV-1 antigen bands were evaluated by logistic regression analysis. A total of 765 HIV-1 infections, 748 (97.8%) with complete test results, were newly diagnosed during the study. A negative or indeterminate HIV antibody assay at diagnosis, symptoms of primary HIV infection, or a negative HIV test during the past 12 mo classified 195 infections (26.1%) as recent (< or = 12 mo). Symptoms of CDC stages B or C classified 161 infections as older (21.5%), and 392 patients with no symptoms remained unclassified. BED-EIA ruled 65% of the 195 recent infections as recent and 80% of the 161 older infections as older. Two INNO-LIA algorithms showed 50% and 40% sensitivity combined with 95% and 99% specificity, respectively. Estimation of recent infection in the entire study population, based on actual results of the three tests and adjusted for a test's sensitivity and specificity, yielded 37% for BED-EIA compared to 35% and 33% for the two INNO-LIA algorithms. Window-based estimation with BED-EIA yielded 41% (95% confidence interval 36%-46%). CONCLUSIONS: Recency information can be extracted from INNO-LIA-based confirmatory testing at no additional costs. This method should improve epidemiologic surveillance in countries that routinely use INNO-LIA for HIV confirmation.

Reference values for peripheral blood lymphocyte phenotypes applicable to the healthy adult population in Switzerland.

OBJECTIVES: Use of domestic reference values is known to improve the accuracy of flow cytometric analysis by integrating local variation due to race, gender, and age. In the absence of previously published estimates, we now report establishment of reference values for a wide range of peripheral blood lymphocyte phenotypes applicable to the healthy adult population in Switzerland and other regions with similar demographic characteristics. METHODS: A representative sample population was recruited from among well characterized local blood donors (n = 70) and quantitative multiparametric flow cytometry used to estimate absolute and proportional values for a range of T-, B-, and NK-cell subsets, including those associated with activation and maturity. Distribution-free methods were then applied to generate 95% reference values and to estimate the significance of gender and age-related differences. RESULTS: Reference values were obtained for the absolute and proportional levels of total CD3(+) T cells (536-1787 cells/microL, 54.90-84.03%), helper CD4(+) T cells (309-1139 cells/microL, 32.53-62.88%), cytotoxic CD8(+) T cells (137-823 cells/microL, 11.55-38.60%), activated CD3(+) T cells expressing CD25 (7-94 cells/microL, 0.50-5.95%), CD38 (102-554 cells/microL, 5.98-26.80%), HLA-DR (18-186 cells/microL, 1.25-9.68%) or CD38/HLA-DR (4-52 cells/microL, 0.30-2.30%), activated CD4(+) T cells expressing CD25 (7-52 cells/microL, 0.33-2.80%), CD38 (69-547 cells/microL, 6.13-32.20%), HLA-DR (11-55 cells/microL, 0.80-4.43%) or CD38/HLA-DR (4-22 cells/microL, 0.30-1.35%), activated CD8(+) T cells expressing CD25 (0-12 cells/microL, 0.00-0.69%), CD38 (13-124 cells/microL, 0.93-7.03%), HLA-DR (6-108 cells/microL, 0.33-6.38%) or CD38/HLA-DR (2-47 cells/microL, 0.13-2.68%), naive CD4(+) T cells expressing CD45RA(+)/CD62L(+) (84-761 cells/microL, 9.48-41.88%), naive CD8(+) T cells expressing CD45RA(+)/CD62L(+) (42-360 cells/microL, 3.68-19.23%), memory CD4(+) T cells expressing CD45RO(+) (247-807 cells/microL, 16.50-42.15%), memory CD8(+) T cells expressing CD45RO(+) (72-377 cells/microL, 3.78-22.80%), B-cells expressing CD19 (72-460 cells/microL, 4.70-19.13%) or CD20 (66-529 cells/microL, 4.63-21.00%), total CD3(-)/(CD16(+)/CD56(+)) NK-cells (77-427 cells/microL, 5.35-30.93%), and activated NK-cells expressing CD25 (0-10 cells/microL, 0-0.50%) or HLA-DR (3-99 cells/microL, 0.20-7.28%). CONCLUSION: It is anticipated that availability of localized reference values for an extended range of peripheral blood lymphocyte phenotypes should supplement previously published reference values and enhance the utility of flow cytometric analysis undertaken in Switzerland.