Adenovirus expressing a bioluminescence reporter gene and cMAGI cell assay for the detection of HIV-1.
We report a fast, highly sensitive method for detecting and testing drug resistance of M-tropic and T-tropic laboratory and primary HIV-1 isolates. cMAGI cells are infected with an adenovirus vector harboring the luciferase reporter gene controlled by HIV-1 Tat-responsive element, TAR. HIV-1 Tat production by HIV-1 chronically infected cells, or by cMAGI cells as early as two days after being acutely infected with HIV-1, is readily monitored in the presence or absence of antiviral drugs. This method is more sensitive than HIV-1 Tat dependant production of beta-galactosidase in the cMAGI cells. The fast answer, ease and sensitivity as well as the possibility of using this method in high throughput screening, makes it an very attractive tool for phenotypic detection of HIV-1 in clinical samples as well as a sensitive assay for monitoring drug resistant HIV-1 variants. This method can also be used for discovery of novel anti HIV-1 drugs.
Ruth Ben-Ari Institute of Clinical Immunology & AIDS Center, Kaplan Medical Center, Hebrew University Hadassah Medical School, Rehovot 76100, Israel. borkow@agri.huji.ac.il
Gadi
G
Borkow
Humberto Herman
HH
Lara
Mila
M
Ayash-Rashkovsky
Einat
E
Tavor
Aviva
A
Lapidot
Zvi
Z
Bentwich
Alik
A
Honigman
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D019380:Anti-HIV Agents / Q000494:pharmacology*; D015496:CD4-Positive T-Lymphocytes / Q000821:virology*; D002460:Cell Line; D005786:Gene Expression Regulation; D015696:Gene Products, tat / Q000378:metabolism; D017930:Genes, Reporter; D005822:Genetic Vectors; D016325:HIV Long Terminal Repeat; D015497:HIV-1 / Q000187:drug effects* / Q000472:pathogenicity; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism*; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008826:Microbial Sensitivity Tests / Q000379:methods; D012680:Sensitivity and Specificity; D013997:Time Factors; D054322:tat Gene Products, Human Immunodeficiency Virus
D019380:Anti-HIV Agents; D015696:Gene Products, tat; D008164:Luminescent Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus; D008156:Luciferases