Patent ID: 7306901
Filing Date: 2007-12-11
Classification: C07K,C12N,C12Q,G01N,G16B,Y02A

Abstract:
1. A method for determining the phenotypic susceptibility of at least one human immunodeficiency virus to at least one drug, comprising: i) obtaining at least one sample comprising human immunodeficiency virus (HIV) RNA, wherein the sample comprises at least one envelope (env) gene or a portion thereof, wherein the portion thereof includes at least gp160, gp120 or gp41; ii) reverse-transcribing and amplifying the HIV RNA, with primers specific for env region of the HIV genome, to obtain at least one amplicon comprising the at least one env gene or a portion thereof, wherein the portion thereof includes at least gp160, gp120 or gp41 wherein at least one primer is selected from SEQ ID NO: 1, 3, 4, 6, 10, 19 and 20; iii) using nucleic acid amplification to generate at least one plasmid containing a wild-type HIV sequence with a deletion in the env region comprising at least 500 bp of the HIV env gene; iv) homologously recombining or ligating the at least one amplicon with the at least one plasmid comprising the wild-type HIV sequence with a deletion in the env gene or a portion thereof, wherein the portion thereof includes at least gp160, gp120 or gp41, to prepare at least one recombinant virus; and v) monitoring the at least one recombinant virus in the presence of the at least one drug to determine the phenotypic susceptibility, wherein the phenotypic susceptibility of at least one human immunodeficiency virus to at least one drug is determined by the replicative capacity of the at least one recombinant virus in the presence of the at least one drug.