Detection of drug resistance mutations in infants' DBS by sequencing and point mutation assays

• Published in: Antiviral therapy Vol. 19; p. A86
• Main Authors: Derache, A, Mutsvangwa, J, Beck, I, Manhanzva, M T, Mtapuri-Zinyowera, S, Gwanzura, L, Frenkel, L, Katzenstein, D
• Format: Journal Article
• Language: English
• Published: 01.01.2014
• Subjects: Human immunodeficiency virus
• ISSN: 1359-6535

HIV-infected infants exposed to nevirapine (NVP) may harbor drug resistance mutations (DRM) in proviral DNA, impacting treatment decisions. HIV amplicon from dried blood spots (DBS) for early infant diagnosis (EID) was assessed by Sanger sequencing (SS), the oligonucleotide ligation assay (OLA) and allele-specific PCR (ASP) to identify DRM among subtype C NVP exposed infants. NVP resistance mutations were detected by OLA in 39% of selected infected infants exposed to NVP which were confirmed in 36/48 (75%) by sequencing. The ASP and OLA showed a good correlation (85%) for K103N and Y181C. Misclassifications were mainly due to mismatches between the template and the primers, highlighting HIV genetic diversity which may limit sensitivity of PMA. DRM >20% by SS and additional NNRTI minority variants were detected only with ASP and OLA among NVP exposed infants. Although the clinical relevance of these DRM is controversial, lower cost PMA could be used to guide therapy among infants infected despite ART.