alexa HIV type-1 drug resistance testing on dried blood spots is feasible and reliable in patients who fail antiretroviral therapy in rural Tanzania.
Immunology

Immunology

Journal of Vaccines & Vaccination

Author(s): Johannessen A, HolbergPetersen M, Lvgaarden G, Naman E, Ormaasen V,

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Abstract BACKGROUND: HIV type-1 (HIV-1) drug resistance testing is rarely available in resource-limited settings because of high costs and stringent requirements for storage and transport of plasma. Dried blood spots (DBS) can be a convenient alternative to plasma, but the use of DBS needs validation under field conditions. We assessed the performance of DBS in genotypic resistance testing of patients who failed first-line antiretroviral therapy (ART) in rural Tanzania. METHODS: A total of 36 ART-experienced patients with viral loads >1,000 copies/ml (median 15,180 copies/ml [range 1,350-3,683,000]) and with various HIV-1 subtypes were selected for resistance testing. DBS were stored with desiccant at ambient temperature for a median of 29 days (range 8-89). Samples were amplified using an in-house reverse transcriptase-nested PCR method and sequenced using the ViroSeqâ„¢ assay (Abbott Molecular, Des Plaines, IL, USA). DBS-derived genotypes were compared with genotypes from plasma. RESULTS: Overall, 34 of 36 (94\%) DBS specimens were successfully genotyped. In the protease region, of 142 polymorphisms found in plasma, 132 (93\%) were also detected in DBS. In the reverse transcriptase region, of 57 clinically relevant mutations present in plasma, 51 (89\%) were also detected in DBS. A total of 30 of 34 (88\%) patients had identical resistance profiles to antiretroviral drugs in plasma and DBS. CONCLUSIONS: Genotyping was successful in the vast majority of DBS specimens stored at ambient temperature for up to 3 months, and there was high concordance between mutations found in DBS and plasma. Our study suggests that DBS can be a feasible and reliable tool to monitor HIV-1 drug resistance in patients on ART in resource-limited settings. This article was published in Antivir Ther and referenced in Journal of Vaccines & Vaccination

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