Methods to determine HIV-1 ex vivo fitness.
The fitness of human immunodeficiency virus type-1 (HIV-1) appears to be an important piece of the puzzle in understanding the global HIV-1 epidemic and disease progression in infected individuals. We have developed a dual infection/competition assay followed by a sensitive heteroduplex tracking assay (HTA) to measure the relative fitness of any HIV-1 isolate. Differences in fitness between wild-type and drug-resistant HIV-1 isolates can also be used as indicators of the emergence of drug-resistant isolates. This chapter describes the methods utilized to measure viral fitness (relative replicative capacity) during growth competition experiments. HTA is used to evaluate the production of HIV-1 variants in a competition, which can then be used to estimate ex vivo viral fitness.
Abraha
Awet
A
Division of Infectious Diseases, Department of Medicine, Case Western Reserve University, Cleveland, OH, USA.
Troyer
Ryan M
RM
Quiñones-Mateu
Miguel E
ME
Arts
Eric J
EJ
D015752:Genes, env; D015497:HIV-1 / Q000235:genetics / Q000254:growth & development / Q000472:pathogenicity / Q000502:physiology*; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000821:virology; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D014779:Virus Replication / Q000235:genetics / Q000502:physiology*
D009692:Nucleic Acid Heteroduplexes