title: A radiolabeled oligonucleotide ligation assay demonstrates the high frequency of nevirapine resistance mutations in HIV type 1 quasispecies of NVP-treated and untreated mother-infant pairs from Uganda.
abstract: This study explores the levels of NVP and AZT resistance mutations in untreated, NVP- or AZT-treated mother-infant pairs in Uganda. PCR-amplified reverse transcriptase (RT) gene fragments derived from PBMC samples of 85 mothers (10 AZT treated, 35 NVP treated, and 40 untreated) and their 52 infected infants (5 AZT, 9 NVP, and 38 untreated) were classified as subtype A (59%), D (29%), C (3%), and recombinant forms (9%) by population sequencing. Only 16% of the NVP-treated infected mothers and infants harbored either the K103N or the Y181C at 6 weeks postdelivery. The majority of these samples (n = 107) were then analyzed using a radiolabeled oligonucleotide ligation assay (OLA) specific for K70R, K103N, and Y181C, using nonstandard bases to accommodate sequence heterogeneity. By OLA, 43% of the NVP-treated group had K103N and/or Y181C mutations in their HIV-1 population, using >0.6% cutoff based on a comparative clonal analysis of clinical isolates. Surprisingly, an equal fraction of the untreated and NVP-treated mother-infant group had the K103N mutation in their HIV-1 population in the range of 0.6-5%. These findings suggest a relatively high frequency of K103N mutation in the drug-naive, subtype A and D infected Ugandan population as compared to the very low frequency of the Y181C and K70R mutation (<0.6%). The prevalence of the K103N mutations may be related to its low fitness cost and high genetic stability. The persistence of these mutations may reduce the effectiveness of subsequent NVP use in treatment or prevention of perinatal transmission.
authors.affiliation: Division of Infectious Diseases, Department of Medicine, Case Western Reserve University, Cleveland, Ohio 44106, USA.
authors.forename: Ryan M
authors.initials: RM
authors.lastname: Troyer
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Lalonde
authors.forename: Erika
authors.initials: E
authors.lastname: Fraundorf
authors.forename: Korey R
authors.initials: KR
authors.lastname: Demers
authors.forename: Fred
authors.initials: F
authors.lastname: Kyeyune
authors.forename: Peter
authors.initials: P
authors.lastname: Mugyenyi
authors.forename: Aslam
authors.initials: A
authors.lastname: Syed
authors.forename: Christopher C
authors.initials: CC
authors.lastname: Whalen
authors.forename: Francis
authors.initials: F
authors.lastname: Bajunirwe
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Arts
mesh_terms: D019943:Amino Acid Substitution / Q000235:genetics; D019380:Anti-HIV Agents / Q000494:pharmacology*; D016000:Cluster Analysis; D004279:DNA, Viral / Q000235:genetics; D024882:Drug Resistance, Viral; D005821:Genetic Techniques; D015658:HIV Infections / Q000821:virology*; D054303:HIV Reverse Transcriptase / Q000235:genetics; D015497:HIV-1 / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007963:Leukocytes, Mononuclear / Q000821:virology; D009035:Mothers; D020125:Mutation, Missense; D019829:Nevirapine / Q000494:pharmacology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D011995:Recombination, Genetic; D017422:Sequence Analysis, DNA; D014454:Uganda; D015215:Zidovudine / Q000494:pharmacology
chemical_list: D019380:Anti-HIV Agents; D004279:DNA, Viral; D015215:Zidovudine; D019829:Nevirapine; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase