Comparison of methods for in-house screening of HLA-B*57:01 to prevent abacavir hypersensitivity in HIV-1 care.

• Published in: PLoS ONE
• Main Authors: Ward De Spiegelaere, Jan Philippé, Karen Vervisch, Chris Verhofstede, Eva Malatinkova, Maja Kiselinova, Wim Trypsteen, Pawel Bonczkowski, Dirk Vogelaers, Steven Callens, Jean Ruelle, Kabamba Kabeya, Stephane De Wit, Petra Van Acker, Vicky Van Sandt, Marie-Paule Emonds, Paul Coucke, Erica Sermijn, Linos Vandekerckhove
• Format: Article
• Language: English
• Published: Public Library of Science (PLoS) 2015-01-01
• Online Access: http://europepmc.org/articles/PMC4398410?pdf=render

Abacavir is a nucleoside reverse transcriptase inhibitor used as part of combination antiretroviral therapy in HIV-1-infected patients. Because this drug can cause a hypersensitivity reaction that is correlated with the presence of the HLA-B*57:01 allotype, screening for the presence of HLA-B*57:01 is recommended before abacavir initiation. Different genetic assays have been developed for HLA-B*57:01 screening, each with specific sensitivity, turnaround time and assay costs. Here, a new real-time PCR (qPCR) based analysis is described and compared to sequence specific primer PCR with capillary electrophoresis (SSP PCR CE) on 149 patient-derived samples, using sequence specific oligonucleotide hybridization combined with high resolution SSP PCR as gold standard. In addition to these PCR based methods, a complementary approach was developed using flow cytometry with an HLA-B17 specific monoclonal antibody as a pre-screening assay to diminish the number of samples for genetic testing. All three assays had a maximum sensitivity of >99. However, differences in specificity were recorded, i.e. 84.3%, 97.2% and >99% for flow cytometry, qPCR and SSP PCR CE respectively. Our data indicate that the most specific and sensitive of the compared methods is the SSP PCR CE. Flow cytometry pre-screening can substantially decrease the number of genetic tests for HLA-B*57:01 typing in a clinical setting.