New method of analysis of the characteristics of the virus phenotypic of the human immunodeficiency (hiv)

摘要

Analyzing (M) 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, is new. M 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 any of the proteins gag, pol, RNase H, integrase, vif, vpr, tat, rev, vpu, nef, cis-active sequences, long terminal repeats, dimerization sequences, splice-regulating sequences or Rev response elements, all 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 optionally containing a marker gene (MG) activated only after viral infection; and (g) detecting and/or quantifying MG expression in the second cells. An Independent claim is also included for a kit for carrying out M.