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

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

doi:10.1371/journal.pone.0123525

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

PLoS One. 2015 Apr 15;10(4):e0123525. doi: 10.1371/journal.pone.0123525. eCollection 2015.

Authors

Ward De Spiegelaere¹, Jan Philippé², Karen Vervisch¹, Chris Verhofstede², Eva Malatinkova¹, Maja Kiselinova¹, Wim Trypsteen¹, Pawel Bonczkowski¹, Dirk Vogelaers¹, Steven Callens¹, Jean Ruelle³, Kabamba Kabeya⁴, Stephane De Wit⁴, Petra Van Acker⁵, Vicky Van Sandt⁶, Marie-Paule Emonds⁶, Paul Coucke⁵, Erica Sermijn¹, Linos Vandekerckhove¹

Affiliations

¹ Ghent University, Department of Internal Medicine, Ghent, Belgium.
² Ghent University, Department of Clinical Chemistry, Microbiology and Immunology, Ghent, Belgium.
³ Université Catholique de Louvain, IREC, AIDS Reference Laboratory, Brussels, Belgium.
⁴ Saint-Pierre University Hospital, Department of Infectious Diseases, Bruxelles, Belgium.
⁵ Ghent University, Center for Medical Genetics, Ghent, Belgium.
⁶ HILA, Laboratory for Histocompatibility & Immunogenetics Red Cross Flanders, Mechelen, Belgium.

Abstract

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.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Alleles
Antiretroviral Therapy, Highly Active
DNA Primers / chemical synthesis
DNA Primers / genetics
Dideoxynucleosides / administration & dosage*
Dideoxynucleosides / adverse effects
Drug Hypersensitivity / etiology
Drug Hypersensitivity / genetics
Drug Hypersensitivity / immunology
Drug Hypersensitivity / prevention & control*
Electrophoresis, Capillary
Flow Cytometry
Gene Expression
Genetic Testing / methods
HIV Infections / drug therapy*
HIV Infections / genetics
HIV Infections / immunology
HIV Infections / virology
HIV-1 / drug effects
HIV-1 / enzymology
HIV-1 / genetics
HLA-B Antigens / genetics*
HLA-B Antigens / immunology
Humans
Nucleic Acid Hybridization / methods
Real-Time Polymerase Chain Reaction / methods*
Reverse Transcriptase Inhibitors / administration & dosage*
Reverse Transcriptase Inhibitors / adverse effects
Sensitivity and Specificity

Substances

DNA Primers
Dideoxynucleosides
HLA-B Antigens
HLA-B*57:01 antigen
HLA-B17 antigen
Reverse Transcriptase Inhibitors
abacavir

Grants and funding

This work was supported by the Research Fund – Flanders (FWO, 1.8.020.09.N.00 to Linos Vandekerckhove), by the Agency for Innovation by Science and Technology in Flanders (IWT, Grant nr: 111286 and 111393 to Eva Malatinkova and Pawel Bonczkowski), and by the Special Research Grant of Ghent University (BOF, 01N02712 to Maja Kiselinova). Viiv Healthcare and Beckton Dickinson provided financial support for sampling and consumables. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.