Analyzing phenotypic characteristics of human immune deficiency virus, useful for assessing susceptibility to hydroxyurea of virus with mutated protease gene

摘要

Analyzing a phenotypic characteristic (A) of HIV in a patient sample where (A) is the result of one or more mutations in the viral genome that can affect viral infection, and the infectivity of the virus is determined, is new. Analyzing a phenotypic characteristic (A) of HIV in a patient sample where (A) is the result of one or more mutations in the viral genome that can affect viral infection, and the infectivity of the virus is determined, is new. The method comprises: (a) extracting nucleic acid (NA) from the sample; (b) at least one PCR amplification of a segment of NA, using primers that flank at least part of the genome that encodes a protease (Pr), potentially mutated; (c) preparing a vector containing all parts of the HIV genome required for replication except for the segment amplified in (b), optionally also the gene encoding the envelope protein; (d) transfection of a first host cell with this vector and the product of (b), optionally also with a vector that provides the envelope gene, if not already present in the first vector; (e) culturing the cells to produce particles of chimeric virus formed by homologous recombination in a single cycle of replication; (f) using these particles to infect second host cells, able to be infected by (pseudotyped) HIV and containing a marker gene (MG) activated only after viral infection, and (g) comparing MG expression between second cells infected with particles of (f) and those infected with reference particles produced similarly from a sample containing a reference virus. An Independent claim is also included for a kit for the process.