Analyzing phenotype of human immune deficiency virus, useful for optimizing therapy, by cloning segment into viral particle and transfecting cell containing inducible marker gene

摘要

Analyzing phenotype of HIV (human immune deficiency virus), resulting from one or more mutations in the viral genome that influence infection, in a patient sample, is new. Analyzing phenotype of HIV (human immune deficiency virus), resulting from one or more mutations in the viral genome that influence infection, in a patient sample, is new. Nucleic acids are extracted from the sample, segments of them amplified by PCR (polymerase chain reaction) using pairs of primers that flank a genomic sequence susceptible to mutation, and a first host cell (HC1) transfected with: (a) the amplicon; (b) a vector containing parts of the HIV genome required for replication, except for the amplified segment and optionally also the env gene; and (c) if the vector of (b) lacks the env gene, also a second vector containing this gene. Homologous recombination occurs to produce a chimeric virus and HC1 are cultured to produce viral particles (VP) during a single cycle of replication. VP are used to infect at least one second host cell (HC2) that contains a marker gene (MG) that is activated only after viral infection, then the expressed marker detected and/or quantified to detect at least one characteristic of the original HIV. An Independent claim is also included for a kit for performing the new process.