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Titolo:
A novel recombinant marker virus assay for comparing the relative fitness of HIV-1 reverse transcriptase variants
Autore:
Lu, J; Kuritzkes, DR;
Indirizzi:
Univ Colorado, Hlth Sci Ctr, Div Infect Dis, Denver, CO 80262 USA Univ Colorado Denver CO USA 80262 r, Div Infect Dis, Denver, CO 80262 USA
Titolo Testata:
JOURNAL OF ACQUIRED IMMUNE DEFICIENCY SYNDROMES
fascicolo: 1, volume: 27, anno: 2001,
pagine: 7 - 13
SICI:
1525-4135(20010501)27:1<7:ANRMVA>2.0.ZU;2-O
Fonte:
ISI
Lingua:
ENG
Soggetto:
HIGH-LEVEL RESISTANCE; IN-VITRO; ZIDOVUDINE RESISTANCE; PROTEASE INHIBITORS; REPLICATIVE FITNESS; TYPE-1; MUTATION; THERAPY; MUTANTS; GENOME;
Keywords:
fitness; HIV-1; polymerase chain reaction; viral;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
21
Recensione:
Indirizzi per estratti:
Indirizzo: Kuritzkes, DR Univ Colorado, Hlth Sci Ctr, Div Infect Dis, 4200 E 9th Ave,B-168, Denver,CO 80262 USA Univ Colorado 4200 E 9th Ave,B-168 Denver CO USA80262 2 USA
Citazione:
J. Lu e D.R. Kuritzkes, "A novel recombinant marker virus assay for comparing the relative fitness of HIV-1 reverse transcriptase variants", J ACQ IMM D, 27(1), 2001, pp. 7-13

Abstract

A novel recombinant marker virus assay (RMVA) has been developed to perform growth competition assays for assessing fitness of HIV-1. This assay allowed the generation of replication-competent viruses by homologous recombination of polymerase chain reaction (PCR)-derived reverse transcriptase (RT) coding sequences in RT-deleted proviral clones of HIV-1 in which the nef gene was replaced by the Salmonella typhimurium histidinol dehydrogenase (hisD) or human heat-stable placental alkaline phosphatase (PLAP) gene (pHIV Delta RTBalI Delta nefhisD and pHIV Delta RTBalI Delta nefPLAP, respectively). The proportion of a given RT species in a mixed culture was determined byquantifying the linked hisD or FLAP marker gene using real-time PCR. The RMVA was tested by comparing the relative fitness of wild-type and lamivudine-resistant recombinant viruses. The RMVA reproducibly detected differencesin the fitness of these two viruses in growth competition assays. With appropriate modification of the recombination vectors, the RMVA should be useful for analyzing the fitness of viruses resistant to protease, integrase, or fusion inhibitors and should be applicable in clinical research.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 04/04/20 alle ore 21:54:39