title: Pyrosequencing dried blood spots reveals differences in HIV drug resistance between treatment naïve and experienced patients.
abstract: Dried blood spots (DBS) are an alternative specimen collection format for HIV-1 genotyping. DBS produce HIV genotyping results that are robust and equivalent to plasma when using conventional sequencing methods. However, using tagged, pooled pyrosequencing, we demonstrate that concordance between plasma and DBS is not absolute and varies according to viral load (VL), duration of HIV infection and antiretroviral therapy (ART) status. The plasma/DBS concordance is the highest when VL is ≥5,000 copies/ml and/or the patient has no ART exposure and/or when the duration of HIV infection is ≤2 years. Stepwise regression analysis revealed that VL is most important independent predictor for concordance of DBS with plasma genotypes. This is the first study to use next generation sequencing to identify discordance between DBS and plasma genotypes. Consideration should be given to VL, duration of infection, and ART exposure when interpreting DBS genotypes produced using next generation sequencing. These findings are of particular significance when DBS are to be used for clinical monitoring purposes.
authors: National HIV & Retrovirology Laboratories, National Microbiology Laboratory, Public Health Agency of Canada, Ottawa, Canada.
authors: Hezhao
authors: H
authors: Ji
authors: Yang
authors: Y
authors: Li
authors: Binhua
authors: B
authors: Liang
authors: Richard
authors: R
authors: Pilon
authors: Paul
authors: P
authors: MacPherson
authors: Michèle
authors: M
authors: Bergeron
authors: John
authors: J
authors: Kim
authors: Morag
authors: M
authors: Graham
authors: Gary
authors: G
authors: Van Domselaar
authors: Paul
authors: P
authors: Sandstrom
authors: James
authors: J
authors: Brooks
mesh_terms: D000293:Adolescent; D000328:Adult; D019380:Anti-HIV Agents / Q000494:pharmacology* / Q000627:therapeutic use; D059788:Dried Blood Spot Testing / Q000379:methods*; D024882:Drug Resistance, Viral / Q000235:genetics*; D005260:Female; D060005:Genotyping Techniques; D015658:HIV Infections / Q000097:blood* / Q000188:drug therapy; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics* / Q000502:physiology*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000821:virology; D008297:Male; D008875:Middle Aged; D010949:Plasma / Q000187:drug effects / Q000821:virology; D017422:Sequence Analysis, DNA / Q000379:methods*; D019562:Viral Load / Q000187:drug effects; D055815:Young Adult
chemical_list: D019380:Anti-HIV Agents