Advanced search

Line probe assay for rapid detection of drug-selected mutations in the human immunodeficiency virus type 1 reverse transcriptase gene.

Author
Organization
Abstract
Upon prolonged treatment with various antiretroviral nucleoside analogs such as 3'-azido-3'-deoxythymidine, 2',3'-dideoxyinosine, 2',3'-dideoxycytidine, (-)-beta-L-2',3'-dideoxy-3'-thiacytidine, and 2',3'-didehydro-3'-deoxythymidine, selection of human immunodeficiency virus type 1 (HIV-1) strains with mutations in the reverse transcriptase (RT) gene has been reported. We designed a reverse hybridization line probe assay (LiPA) for the rapid and simultaneous characterization of the following variations in the RT gene: M41 or WI; T69, N69, A69, or D69; K70 or R70; L74 or V74; V75 or T75; M184, I184, or V184; T215, Y215, or F215; and K219, Q219, or E219. Nucleotide polymorphisms for codon L41 (TTG or CTG), T69 (ACT or ACA), V75 (GTA or GTG), T215 (ACC or ACT), and Y215 (TAG or TAT) could be detected. In addition to the codons mentioned above, several third-letter polymorphisms in the direct vicinity of the target codons (E40, E42, K43, K73, D76, Q182, Y183, D185, G213, F214, and L214) were found, and specific probes were selected. In total, 48 probes were designed and applied on the LiPA test strips and optimized with a well-characterized and representative reference panel. Plasma samples from 358 HIV-infected patients were analyzed with all 48 probes. The amino acid profiles could be deduced by LiPA hybridization in an average of 92.7% of the samples for each individual codon. When combined with changes in viral load and CD4(+) T-tell count, this LiPA approach proved to be useful in studying genetic resistance in follow-up samples from antiretroviral agent-treated HIV-1-infected individuals.

Citation

Please use this url to cite or link to this publication:

Chicago
STUYVER, L, A WYSEUR, A ROMBOUT, J LOUWAGIE, T SCARCEZ, Chris Verhofstede, D RIMLAND, RF SCHINAZI, and R ROSSAU. 1997. “Line Probe Assay for Rapid Detection of Drug-selected Mutations in the Human Immunodeficiency Virus Type 1 Reverse Transcriptase Gene.” Antimicrobial Agents and Chemotherapy 41 (2): 284–291.
APA
STUYVER, L., WYSEUR, A., ROMBOUT, A., LOUWAGIE, J., SCARCEZ, T., Verhofstede, C., RIMLAND, D., et al. (1997). Line probe assay for rapid detection of drug-selected mutations in the human immunodeficiency virus type 1 reverse transcriptase gene. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY, 41(2), 284–291.
Vancouver
1.
STUYVER L, WYSEUR A, ROMBOUT A, LOUWAGIE J, SCARCEZ T, Verhofstede C, et al. Line probe assay for rapid detection of drug-selected mutations in the human immunodeficiency virus type 1 reverse transcriptase gene. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY. 1997;41(2):284–91.
MLA
STUYVER, L, A WYSEUR, A ROMBOUT, et al. “Line Probe Assay for Rapid Detection of Drug-selected Mutations in the Human Immunodeficiency Virus Type 1 Reverse Transcriptase Gene.” ANTIMICROBIAL AGENTS AND CHEMOTHERAPY 41.2 (1997): 284–291. Print.
@article{181987,
  abstract     = {Upon prolonged treatment with various antiretroviral nucleoside analogs such as 3'-azido-3'-deoxythymidine, 2',3'-dideoxyinosine, 2',3'-dideoxycytidine, (-)-beta-L-2',3'-dideoxy-3'-thiacytidine, and 2',3'-didehydro-3'-deoxythymidine, selection of human immunodeficiency virus type 1 (HIV-1) strains with mutations in the reverse transcriptase (RT) gene has been reported. We designed a reverse hybridization line probe assay (LiPA) for the rapid and simultaneous characterization of the following variations in the RT gene: M41 or WI; T69, N69, A69, or D69; K70 or R70; L74 or V74; V75 or T75; M184, I184, or V184; T215, Y215, or F215; and K219, Q219, or E219. Nucleotide polymorphisms for codon L41 (TTG or CTG), T69 (ACT or ACA), V75 (GTA or GTG), T215 (ACC or ACT), and Y215 (TAG or TAT) could be detected. In addition to the codons mentioned above, several third-letter polymorphisms in the direct vicinity of the target codons (E40, E42, K43, K73, D76, Q182, Y183, D185, G213, F214, and L214) were found, and specific probes were selected. In total, 48 probes were designed and applied on the LiPA test strips and optimized with a well-characterized and representative reference panel. Plasma samples from 358 HIV-infected patients were analyzed with all 48 probes. The amino acid profiles could be deduced by LiPA hybridization in an average of 92.7\% of the samples for each individual codon. When combined with changes in viral load and CD4(+) T-tell count, this LiPA approach proved to be useful in studying genetic resistance in follow-up samples from antiretroviral agent-treated HIV-1-infected individuals.},
  author       = {STUYVER, L and WYSEUR, A and ROMBOUT, A and LOUWAGIE, J and SCARCEZ, T and Verhofstede, Chris and RIMLAND, D and SCHINAZI, RF and ROSSAU, R},
  issn         = {0066-4804},
  journal      = {ANTIMICROBIAL AGENTS AND CHEMOTHERAPY},
  language     = {eng},
  number       = {2},
  pages        = {284--291},
  title        = {Line probe assay for rapid detection of drug-selected mutations in the human immunodeficiency virus type 1 reverse transcriptase gene.},
  volume       = {41},
  year         = {1997},
}