Factors affecting relative fitness measurements in pairwise competition assays of human immunodeficiency viruses
Autor: | Dylan H. Westfall, James I. Mullins, Robert A. Smith, Moon S. Kim, J. Victor Swain, Siriphan Manocheewa, Brandon S. Maust, Erinn Lanxon-Cookson, Morgane Rolland |
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Rok vydání: | 2013 |
Předmět: |
Genetics
Mutation Virus Cultivation media_common.quotation_subject viruses Mutant Drug resistance Biology medicine.disease_cause Virus Replication Competition (biology) Article Multiplicity of infection Viral replication Cell culture Virology Drug Resistance Viral medicine HIV-1 Leukocytes Mononuclear Humans Pairwise comparison Cells Cultured media_common |
Zdroj: | Journal of virological methods. 194(1-2) |
ISSN: | 1879-0984 |
Popis: | 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), and a multipoint [1 + s 4,7] algorithm that uses data points within the logarithmic phase of viral growth for assessing fitness differences |
Databáze: | OpenAIRE |
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