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Context: The Centers for Disease Control and Prevention (CDC), Division of HIV/AIDS Prevention, spends approximately 50% of its $325 million annual human immunodeficiency virus (HIV) prevention funds for HIV-testing services. An accurate estimate of the costs of HIV testing in various settings is essential for efficient allocation of HIV prevention resources., Objectives: To assess the costs of HIV-testing interventions using different costing methods., Design, Settings, and Participants: We used the microcosting-direct measurement method to assess the costs of HIV-testing interventions in nonclinical settings, and we compared these results with those from 3 other costing methods: microcosting-staff allocation, where the labor cost was derived from the proportion of each staff person's time allocated to HIV testing interventions; gross costing, where the New York State Medicaid payment for HIV testing was used to estimate program costs, and program budget, where the program cost was assumed to be the total funding provided by Centers for Disease Control and Prevention., Main Outcome Measures: Total program cost, cost per person tested, and cost per person notified of new HIV diagnosis., Results: The median costs per person notified of a new HIV diagnosis were $12 475, $15 018, $2697, and $20 144 based on microcosting-direct measurement, microcosting-staff allocation, gross costing, and program budget methods, respectively. Compared with the microcosting-direct measurement method, the cost was 78% lower with gross costing, and 20% and 61% higher using the microcosting-staff allocation and program budget methods, respectively., Conclusions: Our analysis showed that HIV-testing program cost estimates vary widely by costing methods. However, the choice of a particular costing method may depend on the research question being addressed. Although program budget and gross-costing methods may be attractive because of their simplicity, only the microcosting-direct measurement method can identify important determinants of the program costs and provide guidance to improve efficiency.