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Titolo:
Comparison of two commercial assays for the detection of insertion mutations of HIV-1 reverse transcriptase
Autore:
Koch, N; Tamalet, C; Tivoli, N; Fantini, J; Yahi, N;
Indirizzi:
CHRU La Timone, Virol Lab, F-13005 Marseille, France CHRU La Timone Marseille France F-13005 l Lab, F-13005 Marseille, France Fac Sci St Jerome, UMR INRA, Lab Mediterraneen Rech Nutr, F-130013 Marseille, France Fac Sci St Jerome Marseille France F-130013 , F-130013 Marseille, France
Titolo Testata:
JOURNAL OF CLINICAL VIROLOGY
fascicolo: 2, volume: 21, anno: 2001,
pagine: 153 - 162
SICI:
1386-6532(200105)21:2<153:COTCAF>2.0.ZU;2-2
Fonte:
ISI
Lingua:
ENG
Soggetto:
IMMUNODEFICIENCY-VIRUS TYPE-1; LINE PROBE ASSAY; DRUG-RESISTANCE; PROTEASE INHIBITORS; COMBINATION THERAPY; SEQUENCE-ANALYSIS; INDIVIDUALS; POLYMORPHISMS; SAQUINAVIR; GENOTYPES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Citazioni:
22
Recensione:
Indirizzi per estratti:
Indirizzo: Tamalet, C CHRU La Timone, Virol Lab, 264 Rue St Pierre, F-13005 Marseille, France CHRU La Timone 264 Rue St Pierre Marseille France F-13005 ance
Citazione:
N. Koch et al., "Comparison of two commercial assays for the detection of insertion mutations of HIV-1 reverse transcriptase", J CLIN VIRO, 21(2), 2001, pp. 153-162

Abstract

Insertions in the beta3-beta4 fingers subdomain of HIV-1 reverse transcriptase (RT) confer cross-resistance to various nucleoside analogs. The detection of these rearrangements in the region of codons 67-70 of RT is of primary importance for adapting and optimizing combination treatment regimen. Recent reports suggest that some genotyping techniques based on the hybridization of oligonucleotide probes may fail to detect insertion mutants of HIV-1 RT. In the present study, we have evaluated the efficiency of two commercial kits TruGene (based on Dye Primer sequencing) and Viroseq (Big Dye Terminator technique) for the detection of insertion mutations. The data were compared with an in-house dRhodamine sequencing method. Overall, all these cycle sequencing techniques were operative ill the detection of insertion mutants. The best peak homogeneity in the electrophoregrams was observed withthe Dye primer technique. However, specific compression artifacts were frequently encountered with this technique, rendering ambiguous the interpretation of the electrophoregrams in several regions of the sequence. This shortcoming did not occur with dRhodamine Dye terminator or Bigdye terminator cycle sequencing. In any case, a manual inspection of the electrophoregrams is highly recommended, for all types of cycle sequencing techniques, especially for detecting new mutational patterns of the RT and protease genes. Finally, some specific problems were encountered with the softwares provided with both Trugene and Viroseq kits. (C) 2001 Elsevier Science B.V. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 30/03/20 alle ore 00:13:36