Statistical positivity criteria for the analysis of ELISpot assay data in HIV-1 vaccine trials

J Immunol Methods. 2006 Aug 31;315(1-2):121-32. doi: 10.1016/j.jim.2006.07.015. Epub 2006 Aug 15.


The enzyme-linked immunospot (ELISpot) assay is one of the leading technologies used to measure cellular responses in many settings including HIV vaccine clinical trials. HIV-1-specific effector T lymphocyte responses are considered necessary in the control of infection. Accurate measurement and summary of cellular immune responses are critical to HIV research. ELISpot assay readout is often dichotomized into positive/negative responses according to some pre-specified criteria. Given the increase in the number of replicates used for each stimulation with current assay configurations in the HIV Vaccine Trials Network (HVTN) laboratories, a new approach is now possible. We propose an objective criteria based on a hypothesis-driven method that controls the false positive rate while maintaining sensitivity to each of the antigen-specific responses under study. The new approach is compared to other commonly employed criteria using real and simulated data.

Publication types

  • Evaluation Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • AIDS Vaccines / immunology*
  • Clinical Trials as Topic / methods
  • Clinical Trials as Topic / standards*
  • Computer Simulation
  • Data Interpretation, Statistical
  • Enzyme-Linked Immunosorbent Assay / methods
  • Enzyme-Linked Immunosorbent Assay / standards*
  • Enzyme-Linked Immunosorbent Assay / statistics & numerical data
  • False Positive Reactions
  • HIV Seropositivity / immunology*
  • HIV Seropositivity / therapy
  • HIV-1 / immunology*
  • Humans
  • Sensitivity and Specificity
  • T-Lymphocytes / immunology*


  • AIDS Vaccines