title: Evaluation of dried blood spots collected on filter papers from three manufacturers stored at ambient temperature for application in HIV-1 drug resistance monitoring.
abstract: As more HIV-infected people gain access to antiretroviral therapy (ART), monitoring HIV drug resistance (HIVDR) becomes essential to combat both acquired and transmitted HIVDR. Studies have demonstrated dried blood spots (DBS) are a suitable alternative in HIVDR monitoring using DBS collected on Whatman 903 (W-903). In this study, we sought to evaluate two other commercially available filter papers, Ahlstrom 226 (A-226) and Munktell TFN (M-TFN), for HIVDR genotyping following ambient temperature storage. DBS were prepared from remnant blood specimens collected from 334 ART patients and stored at ambient temperature for a median time of 30 days. HIV-1 viral load was determined using NucliSENS EasyQ® HIV-1 v2.0 RUO test kits prior to genotyping of the protease and reverse transcriptase regions of the HIV-1 pol gene using an in-house assay. Among the DBS tested, 26 specimens had a viral load ≥ 1000 copies/mL in all three types of filter paper and were included in the genotyping analysis. Genotyping efficiencies were similar between DBS collected on W-903 (92.3%), A-226 (88.5%), and M-TFN (92.3%) filter papers (P = 1.00). We identified 50 DR-associated mutations in DBS collected on W-903, 33 in DBS collected on A-226, and 48 in DBS collected on M-TFN, resulting in mutation detection sensitivities of 66.0% for A-226 and 88.0% for M-TFN when compared to W-903. Our data indicate that differences among filter papers may exist at this storage condition and warrant further studies evaluating filter paper type for HIVDR monitoring.
authors.affiliation: International Laboratory Branch, Division of Global HIV/AIDS, Center for Global Health, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.
authors.forename: Erin K
authors.initials: EK
authors.lastname: Rottinghaus
authors.affiliation: International Laboratory Branch, Division of Global HIV/AIDS, Center for Global Health, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.
authors.forename: R Suzanne
authors.initials: RS
authors.lastname: Beard
authors.affiliation: CDC-Botswana, Gaborone, Botswana.
authors.forename: Ebi
authors.initials: E
authors.lastname: Bile
authors.affiliation: CDC-Botswana, Gaborone, Botswana.
authors.forename: Mosetsanagape
authors.initials: M
authors.lastname: Modukanele
authors.affiliation: CDC-Botswana, Gaborone, Botswana.
authors.forename: Maruping
authors.initials: M
authors.lastname: Maruping
authors.affiliation: Botswana Ministry of Health, Gaborone, Botswana.
authors.forename: Madisa
authors.initials: M
authors.lastname: Mine
authors.affiliation: International Laboratory Branch, Division of Global HIV/AIDS, Center for Global Health, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.
authors.forename: John
authors.initials: J
authors.lastname: Nkengasong
authors.affiliation: International Laboratory Branch, Division of Global HIV/AIDS, Center for Global Health, Centers for Disease Control and Prevention, Atlanta, Georgia, United States of America.
authors.forename: Chunfu
authors.initials: C
authors.lastname: Yang
mesh_terms: D059788:Dried Blood Spot Testing / Q000379:methods*; D024882:Drug Resistance, Viral; D005374:Filtration / Q000379:methods; D005838:Genotype; D015658:HIV Infections / Q000097:blood* / Q000188:drug therapy; D015497:HIV-1 / Q000235:genetics* / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D013696:Temperature; D019562:Viral Load