Evaluation of dried blood spots collected on filter papers from three manufacturers stored at ambient temperature for application in HIV-1 drug resistance monitoring

• Published in: PloS one Vol. 9; no. 10; p. e109060
• Main Authors: Rottinghaus, Erin K, Beard, R Suzanne, Bile, Ebi, Modukanele, Mosetsanagape, Maruping, Maruping, Mine, Madisa, Nkengasong, John, Yang, Chunfu
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 10.10.2014; Public Library of Science (PLoS)
• Subjects: Acquired immune deficiency syndrome; AIDS; Ambient temperature; Antiretroviral agents; Antiretroviral drugs; Antiretroviral therapy; Biology and Life Sciences; Blood; Data processing; Disease control; DNA polymerases; Dried Blood Spot Testing - methods; Drug resistance; Drug Resistance, Viral; Drug therapy; Filter paper; Filtration - methods; Genotype; Genotyping; Highly active antiretroviral therapy; HIV; HIV Infections - blood; HIV Infections - drug therapy; HIV-1 - genetics; HIV-1 - isolation & purification; Human immunodeficiency virus; Humans; Medicine and health sciences; Molecular Sequence Data; Monitoring; Mutation; Pol gene; Proteases; RNA-directed DNA polymerase; Temperature; Temperature effects; Viral Load; Atlanta Georgia; Georgia; United States > US; Botswana
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0109060

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.