HIV-1 drug resistance testing from dried blood spots collected in rural Tanzania using the ViroSeq HIV-1 Genotyping System

Objectives: 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. Patients and 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 -20C 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 10000copies/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 <10000 copies/mL. In DBS that yielded a genotype there was high concordance with a plasma-based assay.