Soussan P, Collin G, Descamps D, Foiny P, Brun-Vezinet F; Conference on Retroviruses and Opportunistic Infections.
Program Abstr 4th Conf Retrovir Oppor Infect Conf Retrovir Oppor Infect 4th 1997 Wash DC. 1997 Jan 22-26; 4th: 182 (abstract no. 634).
Hopital Bichat-Claude Bernard, Paris, France.
Methionine 184 Valine (M184V) mutation emerges in patients receiving lamivudine (3TC) therapy. Selective PCR for detection of this mutation generated unreliable results. M184V mutation could be associated with phenotypic reversion of zidovudine (ZDV) resistance. Objective: To develop a new rapid assay for detection of M184V mutation. Methods: Samples were collected from 11 experienced (ZDV+ddI or ZDV+ddC) patients treated by 3TC and ZDV for a 10 month period. We analysed 36 PBMC viral isolates and 22 plasma specimens from these patients. RT gene from codon 154 to 261 was amplified by nested PCR from proviral DNA infected PBMC and plasma specimens. RFDA were performed using 2 restriction enzymes Nla III (recognition site 5' ACATG/ 3') and BsaA I (recognition site 5' AC/GTG 3') allowing determination, on agarose gel electrophoresis, of 184M wild-type (ATG) or 184V mutant (GTG) strains. RFDA results were confirmed by sequence analyses. Phenotypic susceptibility to 3TC of these isolates was determined. Results: Before 3TC therapy, all isolates were only digested by Nla III restriction enzyme and confirmed as wild type by sequence analysis. M184V mutation generated a modification of the 184 restriction site substituting Bsa AI for Nla III. After 4 to 24 weeks of 3TC therapy, this substitution allowed detection of the aminoacid switch M (ATG) to V (GTG) at codon 184 in all patients. As sequencing data, phenotypic assays were in complete agreement with RFDA. Phenotypic reversion of ZDV resistance was analysed. Conclusion: RFDA allowed a rapid and simple determination of M184V mutation and might be worthy of using in patient receiving combination therapy including 3TC and in surveys of the prevalence of primary resistance.
Publication Types:
Keywords:
- Codon
- Didanosine
- HIV-1
- Humans
- Lamivudine
- Mutation
- Zalcitabine
- Zidovudine
- genetics
- reverse transcriptase, Human immunodeficiency virus 1
Other ID:
UI: 102225320
From Meeting Abstracts