Open AccessPyrosequencing Dried Blood Spots Reveals Differences in HIV Drug Resistance between Treatment Naïve and Experienced Patients
Author(s) -
Hezhao Ji,
Li Yang,
Binhua Liang,
Richard Pilon,
Paul MacPherson,
Marcelle Bergeron,
John Kim,
Morag Graham,
Gary Van Domselaar,
Paul Sandstrom,
James Brooks
Publication year - 2013
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0056170
Subject(s) - concordance , genotyping , dried blood spot , pyrosequencing , viral load , human immunodeficiency virus (hiv) , hiv drug resistance , genotype , drug resistance , dried blood , medicine , antiretroviral therapy , biology , virology , immunology , genetics , chemistry , gene , chromatography
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.