Factors affecting relative fitness measurements in pairwise competition assays of human immunodeficiency viruses.

ABSTRACT: Cell culture growth competition assays of human immunodeficiency virus type 1 (HIV-1) are used to estimate viral fitness and quantify the impact of mutations conferring drug resistance and immunological escape. A comprehensive study of growth competition assays was conducted and identified experimental parameters that can impact measurements of relative fitness including multiplicity of infection, viral input ratio, number, timing and interval of time points used to evaluate selective outgrowth, and the algorithm for calculating fitness values. An optimized protocol is developed here that is a multi-point growth competition assay that resolves reproducibly small differences in viral fitness. The optimized protocol uses an MOI of 0.005, a consistent ratio of mutant: parental viruses (70:30)

SUBMITTER: Lanxon-Cookson EC 

PROVIDER: S-EPMC3825816 | biostudies-literature | 2013 Dec

REPOSITORIES: biostudies-literature