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To determine limitations and strengths of three methodologies developed to estimate HIV prevalence and the number of people living with HIV/AIDS (PLWHA).The UNAIDS/WHO Workbook method; the Multiparameter Evidence Synthesis (MPES) adopted by the Health Protection Agency; and the UNAIDS/WHO Estimation and Projection Package (EPP) and Spectrum method were used and their applicability and feasibility were assessed. All methods estimate the number infected in mutually exclusive risk groups among 15–70-year-olds.Using data from the Netherlands, the Workbook method estimated 23 969 PLWHA as of January 2008. MPES estimated 21 444 PLWHA, with a 95% credible interval (CrI) of 17 204–28 694. Adult HIV prevalence was estimated at 0.2% (95% CrI 0.15–0.24%) and 40% (95% CrI 25–55%) were undiagnosed. Spectrum applied gender-specific mortality, resulting in a projected estimate of 19 115 PLWHA.Although outcomes differed between the methods, they broadly concurred. An advantage of MPES is that the proportion diagnosed can be estimated by risk group, which is important for policy guidance. However, before MPES can be used on a larger scale, it should be made more easily applicable. If the aim is not only to obtain annual estimates, but also short-term projections, then EPP and Spectrum are more suitable. Research into developing and refining analytical tools, which make use of all available information, is recommended, especially HIV diagnosed cases, as this information is becoming routinely collected in most countries with concentrated HIV epidemics.