Analysis of Drug Resistance Mutations in Whole Blood DNA from HIV-1 Infected Patients by Single Genome and Ultradeep Sequencing Analysis

Publication date: Available online 30 June 2018
Source:Journal of Virological Methods
Author(s): Alma Sotillo, Olalla Sierra, Lorena Martínez-Prats, Francisca Gutiérrez, Sagrario Zurita, Federico Pulido, Rafael Rubio, Rafael Delgado
In HIV-1 infected patients blood CD4+ T lymphocytes could be a valuable target to analyze drug resistance mutations (DRM) selected over the course of the infection. However, detection of viral resistance mutations in cellular DNA by standard genotype resistance techniques (SGRT) is suboptimal.Whole blood DNA (wbDNA) from 12 HIV-1 infected patients on ART was studied by Single Genome Sequencing (SGS) and 8 of them also by Ultradeep pyrosequencing (UDP).Results were compared with contemporary and historical DRM detected in plasma by SGRT during follow up. All the contemporary DRM detected in plasma from the viremic patients were detected by SGS and UDP (20 from 7 patients and 4 from 5 patients respectively). Out of the 67 historical DRM detected in plasma and no longer present at the time of testing, 38 (57%) were detected by SGS in 12 patients and 27 out of 46 (59%) by UDP in 8 patients. Additional DRM never reported in plasma by SGRT were detected by SGS (12 from 8 patients) and UDP (10 from 8 patients).Analysis of wbDNA from HIV-1 infected patients by SGS and UDP provides proof of concept of the value of blood DNA to investigate current and archived DRM in HIV-1 infected patients on ART.