HIV-1 drug resistance testing from dried blood spots collected in rural Tanzania using the ViroSeq HIV-1 Genotyping System.
OBJECTIVE
To assess whether the commercial ViroSeq HIV-1 Genotyping System (Abbott Molecular, Des Plains, IL, USA) can be used in conjunction with dried blood spots (DBS) for clinical monitoring of drug resistance in patients who fail antiretroviral treatment (ART) in rural Tanzania.


METHODS
Patients at Haydom Lutheran Hospital with confirmed treatment failure (viral load >1000 copies/mL) of a first-line ART regimen were selected for resistance testing. DBS were stored with desiccant at -20 °C for a median of 126 days (range 0-203) and shipped at ambient temperature for 20 days. After manual extraction of nucleic acids, the ViroSeq kit was used for amplification and sequencing. DBS-derived genotypes were compared with those of a plasma-based assay.


RESULTS
Seventeen of 36 (47%) DBS specimens were successfully genotyped. Only 2 of 16 (13%) DBS with a viral load <10,000 copies/mL could be amplified, compared with 15 of 20 (75%) DBS with a viral load >10,000 copies/mL (P = 0.001). In samples that yielded a sequence, all 23 clinically significant reverse transcriptase (RT) mutations in plasma were also detected in DBS. One RT mutation was found in DBS only. In the protease region, 77 polymorphisms were found in plasma, of which 70 (91%) were also detected in DBS. Sixteen of 17 (94%) patients had identical resistance profiles to antiretroviral drugs in plasma and DBS.


CONCLUSIONS
The ViroSeq kit performed well in patients with a high viral load, but failed to genotype most DBS with a viral load <10,000 copies/mL. In DBS that yielded a genotype, there was high concordance with a plasma-based assay.
Department of Infectious Diseases, Oslo University Hospital, Ulleval, Oslo, Norway.
Asgeir
A
Johannessen
Carolina
C
Garrido
Natalia
N
Zahonero
Ezra
E
Naman
Carmen
C
de Mendoza
D044966:Anti-Retroviral Agents / Q000627:therapeutic use*; D024882:Drug Resistance, Viral / Q000235:genetics*; D005838:Genotype; D015658:HIV Infections / Q000097:blood / Q000188:drug therapy* / Q000821:virology; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics*; D006801:Humans; D008826:Microbial Sensitivity Tests; D025202:Molecular Diagnostic Techniques; D009154:Mutation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D017423:Sequence Analysis, RNA; D013048:Specimen Handling; D013636:Tanzania; D017211:Treatment Failure; D019562:Viral Load
D044966:Anti-Retroviral Agents